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Development of a Sensitive and Specific Enzyme-Linked Immunosorbent Assay Based on Recombinant Antigens for Rapid Detection of Antibodies against Mycoplasma agalactiae in Sheep▿ †
Fusco, Marisa
Corona, Lucia
Onni, Toniangelo
Marras, Elisabetta
Longheu, Carla
Idini, Graziano
Tola, Sebastiana
Clinical and Vaccine Immunology
We developed a new recombinant enzyme-linked immunosorbent assay (rELISA) for serodiagnosis of contagious agalactia (CA), a disease caused by Mycoplasma agalactiae in sheep and goats. The assay is based on two M. agalactiae surface proteins, namely, P80 and P55. Identification of these immunodominant and common antigens was accomplished by examining the antibody response elicited in sheep during experimental infection and comparing it to the protein expression profiles of 75 M. agalactiae field strains. Our rELISA was tested with 343 sera, collected from sheep with a laboratory-confirmed diagnosis of CA (n = 223) and from healthy animals (n = 120). All sera had previously been tested by Western blotting (WB) for reactivity against M. agalactiae. In addition, our rELISA was compared with a commercial routine ELISA based on inactivated antigens (CHEKiT). Among the 223 samples that were WB positive for M. agalactiae, 209 (93.7%) tested positive for rP80-P55 with our ELISA, whereas only 164 (73.8%) tested positive with the CHEKiT ELISA. Among the 120 samples tested that were WB negative for M. agalactiae, 96.7% were confirmed as negative with our rELISA, while only 75.8% were confirmed as negative with the CHEKiT ELISA. A comparison of the results with receiver operating characteristic curves indicated that the differences observed between our rELISA and the CHEKiT ELISA are statistically significant. The use of recombinant peptides instead of inactivated antigens could significantly improve the discrimination of positive and negative animals, bringing significant advantages in controlling the import/export of live animals and helping in eradication of this economically detrimental disease.
Related Articles
Mycoplasma agalactiae p40 Gene, a Novel Marker for Diagnosis of Contagious Agalactia in Sheep by Real-Time PCR: Assessment of Analytical Performance and In-House Validation Using Naturally Contaminated Milk Samples▿ †
Oravcová, Katarína
López-Enríquez, Lorena
Rodríguez-Lázaro, David
Hernández, Marta
Journal of Clinical Microbiology
We evaluated the capacity of the Mycoplasma agalactiae p40 gene as a diagnostic marker for contagious agalactia in sheep by quantitative real-time PCR. The p40 gene encodes an immunodominant adhesin that plays a key role in cytoadhesion of M. agalactiae. The assay was 100% specific, with an analytical sensitivity of 1 genome equivalent (GE), a quantification that is highly linear (R2 > 0.992) and efficient (PCR efficiency, >0.992) over a 6-log dynamic range, down to 10 GE. We evaluated the capacity of the assay to detect Mycoplasma agalactiae in 797 milk samples (373 raw sheep milk samples from refrigerated tanks of different farms and 424 milk samples from individual sheep of a flock positive for M. agalactiae). In parallel, we also tested the samples by using microbiological isolation coupled with microscopy identification and by a PCR method recommended by the World Organization for Animal Health. While our assay was able to detect 57 (15.28%) positive samples of the 373 milk samples from different farms, identification by microbiological isolation coupled with microscopy detected only 36 (9.65%) samples, and the conventional PCR detected 31 (8.31%) samples. These findings showed that our assay based on the p40 gene is more specific and sensitive for the detection of M. agalactiae in actual natural samples and, thus, can be a promising alternative tool for diagnosis and epidemiological studies of M. agalactiae infection.
Related Articles
Identification of Novel Immunogenic Proteins from Mycoplasma bovis and Establishment of an Indirect ELISA Based on Recombinant E1 Beta Subunit of the Pyruvate Dehydrogenase Complex
Sun, Zhenhong
Fu, Ping
Wei, Kai
Zhang, Haiyan
Zhang, Yuewei
Xu, Jian
Jiang, Fei
Liu, Xu
Xu, Wei
Wu, Wenxue
PLoS ONE
The pathogen Mycoplasma bovis (M. bovis) is a major cause of respiratory disease, mastitis, and arthritis in cattle. Screening the key immunogenic proteins and updating rapid diagnostic techniques are necessary to the prevention and control of M. bovis infection. In this study, 19 highly immunogenic proteins from M. bovis strain PD were identified using 2-dimensional gel electrophoresis, immunoblotting and MALDI-TOF/TOF MS. Of these 19 proteins, pyruvate dehydrogenase E1 component beta subunit (PDHB) showed excellent immune reactivity and repeatability. PDHB was found to be conserved in different M. bovis isolates, as indicated by Western blot analysis. On the basis of these results, a rPDHB-based indirect ELISA (iELISA) was established for the detection of serum antibodies using prokaryotically expressed recombinant PDHB protein as the coating antigen. The specificity analysis result showed that rPDHB-based iELISA did not react with other pathogens assessed in our study except M. agalactiae (which infects sheep and goats). Moreover, 358 serum samples from several disease-affected cattle feedlots were tested using this iELISA system and a commercial kit, which gave positive rates of 50.8% and 39.9%, respectively. The estimated Kappa agreement coefficient between the two methods was 0.783. Notably, 39 positive serum samples that had been missed by the commercial kit were all found to be positive by Western blot analysis. The detection rate of rPDHB-based iELISA was significantly higher than that of the commercial kit at a serum dilution ratio of 1∶5120 to 1∶10,240 (P<0.05). Taken together, these results provide important information regarding the novel immunogenic proteins of M. bovis. The established rPDHB-based iELISA may be suitable for use as a new method of antibody detection in M. bovis.
Related Articles
The indirect hemagglutination test for the detection of antibodies in cattle naturally infected mycoplasmas.
Cho, H J
Ruhnke, H L
Langford, E V
Canadian Journal of Comparative Medicine
Stable mycoplasma antigens for the indirect hemagglutination test (IHA) were prepared employing glutaraldehyde treated sheep erythrocytes sensitized with Mycoplasma agalactiae subsp. bovis and Mycoplasma bovigenitalium antigens. Employing these antigens mycoplasma antibodies were detected in sera from cattle which had mastitic symptoms due to natural infection with either M. agalactiae subsp. bovis or M. bovigenitalium. A total of 200 cows from four herds were examined at varying intervals for the presence of M. agalactiae subsp. bovis and for the detection of antibody using growth inhibition and IHA tests. Mycoplasmas were isolated from 37 animals. Growth inhibiting antibody was detected from 56 of the 200 animals. In the IHA tests, antibody titer greater than or equal to 1:80 were detected in 148 animals, 76 of these having antibody titers greater than or equal to 1:160, while sera of 116 normal control animals had no growth inhibiting antibody and none had IHA antibody titers greater than 1:40. M. bovigenitalium was isolated from the milk of three of 26 animals in a fifth herd during an outbreak of mastitis. Growth inhibiting antibodies were demonstrated in the sera of ten of the 26 animals. However, the IHA test detected antibody titers of greater than or equal to 1:160 in 13 animals and of 1:80 in one of the 26 animals. To determine the specificity of the IHA tests, M. agalactiae subsp. bovis and M. bovigenitalium antigens were reacted with rabbit hyperimmune typing sera produced against 12 species of bovine mycoplasmatales. Homologous antisera showed IHA antibody titers of 1:1280 and 1:2560 against M. agalactiae subsp. bovis and M. bovigenitalium respectively, whereas heterologous antisera showed IHA antibody titers of less than or equal to 1:20. Also eight type-specific bovine antisera were reacted with M agalactiae subsp. bovis and M. bovigenitalium antigens in homologous and heterologous tests. Homoogous reactions showed IHA antibody titers greater than or equal to 1:320, whereas heterologous reactions showed IHA titers of less than or equal to 1:20. This IHA test promises to be useful for the detection of bovine mycoplasma antibodies in sera from cattle infected with M. agalactiae subsp. bovis or M. bovigenitalium. Thes test is sensitive, reproducible and specific and the technique is relatively simple and rapid. The antigens were stable for at least seven months.
Related Articles
A survey of Mycoplasma agalactiae in dairy sheep farms in Spain
Ariza-Miguel, Jaime
Rodríguez-Lázaro, David
Hernández, Marta
BMC Veterinary Research
Contagious Agalactia (CA) is one of the major animal health problems in small ruminants because of its economic significance. Currently, four Mycoplasma spp. have been associated with this syndrome: M. agalactiae, M. mycoides subsp. capri, M. capricolum subsp. capricolum and M. putrefaciens. Their presence has been evaluated in several studies conducted in CA-endemic countries. However, previous Spanish studies have been focused on caprine CA, and there is a knowledge gap regarding which Mycoplasma species are present in sheep flocks from Spain, which has the second highest number of sheep amongst the 27 European Union member states. Consequently, we investigated the presence and geographic distribution of the four CA-causing mycoplasmas in Spanish dairy sheep farms. This is the first time such an investigation has been performed.
Three hundred thirty nine out of 922 sheep flocks were positive for M. agalactiae by real time PCR (36.8%) and 85 by microbiological identification (9.2%). Interestingly, all 597 milk samples assessed for the presence of M. mycoides subsp. capri, M. capricolum subsp. capricolum and M. putrefaciens tested negative. To evaluate the intermittent excretion of the pathogen in milk, we sampled 391 additional farms from 2 to 5 times, resulting that in 26.3% of the cases a previously positive farm tested negative in a later sampling.
M. agalactiae was the only Mycoplasma species detected in the study area showing a high frequency of presence and wide distribution. Therefore, the establishment of a permanent surveillance network is advantageous, as well as the implementation of control and prevention measures to hinder the dissemination of M. agalactiae and to prevent the entrance of other Mycoplasma species.
Mycoplasma agalactiae; Contagious agalactia; Real time PCR; Sheep; Dairy; Spain
Related Articles
Characterization and Analysis of a Stable Serotype-Associated Membrane Protein (P30) of Mycoplasma agalactiae
Fleury, Bénédicte
Bergonier, Dominique
Berthelot, Xavier
Schlatter, Yvonne
Frey, Joachim
Vilei, Edy M.
Journal of Clinical Microbiology
The gene for a 30-kDa immunodominant antigen, P30, of Mycoplasma agalactiae was cloned from type strain PG2 and expressed in Escherichia coli. P30 is encoded on a monocistronic operon determined by two −10 boxes and a possible −35 region constituting the potential promoter, and a transcription termination site. The gene for the 266-amino-acid protein is preceded by a polypurine-rich region designed as the consensus sequence for a ribosome-binding site. Analysis of the amino acid sequence of P30 revealed the presence of a recognition site for a prokaryotic signal peptidase II at amino acid (aa) 24, indicating that P30 is a transmembrane protein. Moreover, Triton X-114 phase partitioning of M. agalactiae PG2 total antigen revealed that P30 is strongly hydrophobic and hence a possible membrane component. Immunoblot analysis using the monospecific polyclonal anti-P30-His serum indicated that P30 is specific to M. agalactiae. Furthermore, PCR amplification with specific primers for p30 and Southern blot analysis revealed the presence of the gene in all M. agalactiae strains tested and its absence in the other mycoplasma species. Among 27 strains of M. agalactiae studied, 20 strains belonging to the common serotypes A to D, including PG2, expressed P30 or part of it as detected by the monospecific polyclonal anti-P30 antibodies. The other seven strains belonging to the rarely isolated serotypes E to H were negative for P30. The p30 gene was sequenced in 15 strains of M. agalactiae, 10 of which expressed P30 or at least part of it and 5 of which did not express P30. The negative strains carried mutations in both −10 boxes of the promoters. These mutations seem to be responsible for the lack of P30 expression in these strains. Analysis of sera from sheep that were experimentally infected with M. agalactiae revealed that P30 induced a strong and persistent immune response which was still very high two months after infection. In contrast, currently used enzyme-linked immunosorbent assay serology gave only low titers.
Related Articles
Mycoplasma agalactiae MAG_5040 is a Mg2+-Dependent, Sugar-Nonspecific SNase Recognised by the Host Humoral Response during Natural Infection
Cacciotto, Carla
Addis, Maria Filippa
Coradduzza, Elisabetta
Carcangiu, Laura
Nuvoli, Anna Maria
Tore, Gessica
Dore, Gian Mario
Pagnozzi, Daniela
Uzzau, Sergio
Chessa, Bernardo
Pittau, Marco
Alberti, Alberto
PLoS ONE
In this study the enzymatic activity of Mycoplasma agalactiae MAG_5040, a magnesium-dependent nuclease homologue to the staphylococcal SNase was characterized and its antigenicity during natural infections was established. A UGA corrected version of MAG_5040, lacking the region encoding the signal peptide, was expressed in Escherichia coli as a GST fusion protein. Recombinant GST-MAG_5040 exhibits nuclease activity similar to typical sugar-nonspecific endo- and exonucleases, with DNA as the preferred substrate and optimal activity in the presence of 20 mM MgCl2 at temperatures ranging from 37 to 45°C. According to in silico analyses, the position of the gene encoding MAG_5040 is consistently located upstream an ABC transporter, in most sequenced mycoplasmas belonging to the Mycoplasma hominis group. In M. agalactiae, MAG_5040 is transcribed in a polycistronic RNA together with the ABC transporter components and with MAG_5030, which is predicted to be a sugar solute binding protein by 3D modeling and homology search. In a natural model of sheep and goats infection, anti-MAG_5040 antibodies were detected up to 9 months post infection. Taking into account its enzymatic activity, MAG_5040 could play a key role in Mycoplasma agalactiae survival into the host, contributing to host pathogenicity. The identification of MAG_5040 opens new perspectives for the development of suitable tools for the control of contagious agalactia in small ruminants.
Related Articles
VNTR analysis reveals unexpected genetic diversity within Mycoplasma agalactiae, the main causative agent of contagious agalactia
McAuliffe, Laura
Churchward, Colin P
Lawes, Joanna R
Loria, Guido
Ayling, Roger D
Nicholas, Robin AJ
BMC Microbiology
Mycoplasma agalactiae is the main cause of contagious agalactia, a serious disease of sheep and goats, which has major clinical and economic impacts. Previous studies of M. agalactiae have shown it to be unusually homogeneous and there are currently no available epidemiological techniques which enable a high degree of strain differentiation.
We have developed variable number tandem repeat (VNTR) analysis using the sequenced genome of the M. agalactiae type strain PG2. The PG2 genome was found to be replete with tandem repeat sequences and 4 were chosen for further analysis. VNTR 5 was located within the hypothetical protein MAG6170 a predicted lipoprotein. VNTR 14 was intergenic between the hypothetical protein MAG3350 and the hypothetical protein MAG3340. VNTR 17 was intergenic between the hypothetical protein MAG4060 and the hypothetical protein MAG4070 and VNTR 19 spanned the 5' end of the pseudogene for a lipoprotein MAG4310 and the 3' end of the hypothetical lipoprotein MAG4320.
We have investigated the genetic diversity of 88 M. agalactiae isolates of wide geographic origin using VNTR analysis and compared it with pulsed field gel electrophoresis (PFGE) and random amplified polymorphic DNA (RAPD) analysis. Simpson's index of diversity was calculated to be 0.324 for PFGE and 0.574 for VNTR analysis. VNTR analysis revealed unexpected diversity within M. agalactiae with 9 different VNTR types discovered. Some correlation was found between geographical origin and the VNTR type of the isolates.
VNTR analysis represents a useful, rapid first-line test for use in molecular epidemiological analysis of M. agalactiae for outbreak tracing and control.
Related Articles
Evaluation of an indigenous ELISA for diagnosis of Johne's disease and its comparison with commercial kits
Singh, S. V.
Singh, A. V.
Singh, P. K.
Sohal, J. S.
Singh, N. P.
Indian Journal of Microbiology
Country lacks sensitive and indigenous diagnostic kits for the screening of goats and sheep against Johne's disease. Therefore an indigenous ELISA kit was developed using protoplasmic antigen from native Mycobacterium avium subspecies paratuberculosis 'Bison Type' strain of goat origin (Kit 1). In the present study, kit 1 and two commercial kits (kit 2 and 3) were evaluated with respect to 'Gold Standard' fecal culture in 71 animals (55 goats and 16 sheep). Kit 1 using indigenous antigen (protoplasmic antigen) was sensitive at very low concentration (0.1 μgm / well) as compared to purified commercial protoplasmic antigen (4 μgm / well) used in kit 2, in the Type 1 reactors (strong positive as positive). Screening of 71 animals by fecal culture detected 38.0% animals (goats-40.0%, sheep-31.2%) as positive (clinical shedders of bacilli) from these farm animals. Of the farm animals located at Central Institute for Research on Goats, herds of goat were endemic whereas, sheep flocks were comparatively resistant to Johne's disease. The 29.5 and 61.9, 15.4 and 57.7 and 4.2 and 14.0% animals (goats and sheep) were in the category of sero-reactors type 1 and 2 of the ELISA kits 1, 2 and 3, respectively. In the type 1 sero-reactors, sensitivity and specificity of kit 1, 2 and 3 was 53.7 and 86.0, 17.8 and 86.0 and 3.5 and 94.7%, respectively. Indigenous ELISA test (kit 1) was significantly superior for the screening of goatherds and sheep flocks against JD as compared to commercial ELISA kits (Kit 2 and 3). In comparison to kit 2 and 3, kit 1 had highest sensitivity, comparable specificity and substantial to nearly perfect proportional agreement (Kappa Scores) with respect to 'Gold standard' fecal culture in goats and sheep. Disease being endemic in herds and flocks screened using ELISA kits, Type I sero-rectors had better correlation with fecal culture in comparison to Type II sero-reactors therefore, used for estimation of sero-prevalence. Newly developed Indigenous ELISA kit was simple, inexpensive, sensitive and reliable for screening of goats and sheep population against Johne's disease. The study reports high prevalence of Johne's disease in farm goatherds and sheep flocks, using sensitive tests (fecal culture and ELISA kit). Results of Type 1 reaction in kit 1 were optimally correlated with culture and were good for estimating the sero-prevalence. For controlling Johne's disease in endemic herds initial removal of the animals in strong positive category (Tyep 1 reactors), may help to remove heavy shedders.
Johne's disease; Mycobacterium avium subsp. paratuberculosis; Protoplasmic antigen; Diagnosis; Fecal culture; ELISA kit
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Serological Testing Versus Other Strategies for Diagnosis of Active Tuberculosis in India: A Cost-Effectiveness Analysis
Dowdy, David W.
Steingart, Karen R.
Pai, Madhukar
PLoS Medicine
This cost-effectiveness study shows that sputum smear microscopy is the most cost-effective test for active tuberculosis (TB) in India, and liquid culture plus microscopy is more cost-effective for TB diagnosis than serological tests.
Undiagnosed and misdiagnosed tuberculosis (TB) drives the epidemic in India. Serological (antibody detection) TB tests are not recommended by any agency, but widely used in many countries, including the Indian private sector. The cost and impact of using serology compared with other diagnostic techniques is unknown.
Methods and Findings
Taking a patient cohort conservatively equal to the annual number of serological tests done in India (1.5 million adults suspected of having active TB), we used decision analysis to estimate costs and effectiveness of sputum smear microscopy (US$3.62 for two smears), microscopy plus automated liquid culture (mycobacterium growth indicator tube [MGIT], US$20/test), and serological testing (anda-tb ELISA, US$20/test). Data on test accuracy and costs were obtained from published literature. We adopted the perspective of the Indian TB control sector and an analysis frame of 1 year. Our primary outcome was the incremental cost per disability-adjusted life year (DALY) averted. We performed one-way sensitivity analysis on all model parameters, with multiway sensitivity analysis on variables to which the model was most sensitive.
If used instead of sputum microscopy, serology generated an estimated 14,000 more TB diagnoses, but also 121,000 more false-positive diagnoses, 102,000 fewer DALYs averted, and 32,000 more secondary TB cases than microscopy, at approximately four times the incremental cost (US$47.5 million versus US$11.9 million). When added to high-quality sputum smears, MGIT culture was estimated to avert 130,000 incremental DALYs at an incremental cost of US$213 per DALY averted. Serology was dominated by (i.e., more costly and less effective than) MGIT culture and remained less economically favorable than sputum smear or TB culture in one-way and multiway sensitivity analyses.
In India, sputum smear microscopy remains the most cost-effective diagnostic test available for active TB; efforts to increase access to quality-assured microscopy should take priority. In areas where high-quality microscopy exists and resources are sufficient, MGIT culture is more cost-effective than serology as an additional diagnostic test for TB. These data informed a recently published World Health Organization policy statement against serological tests.
Please see later in the article for the Editors' Summary
Editors' Summary
Every year, about 2 million people develop tuberculosis in India—a fifth of the global incidence of this highly contagious bacterial infection. Mycobacterium tuberculosis, the bacterium that causes tuberculosis, is spread in airborne droplets when people with the disease cough or sneeze and usually infects the lungs although it can also infect other organs. The characteristic symptoms of tuberculosis are a persistent cough, weight loss, and night sweats. Diagnostic tests for tuberculosis include sputum smear microscopy (microscopic analysis of mucus brought up from the lungs by coughing), culture (growth) of M. tuberculosis from sputum samples in liquid media (using, for example, a commercial product called the mycobacteria growth indicator tube or MGIT), and nucleic acid amplification tests (which detect the bacterium's genome in patient samples) such as the Xpert MTB/RIF system. Tuberculosis can usually be cured by taking several powerful antibiotics daily for at least 6 months.
Why Was This Study Done?
In India, as elsewhere, undiagnosed and misdiagnosed tuberculosis drives the tuberculosis epidemic by increasing the transmission of M. tuberculosis. Unfortunately, sputum smear microscopy, the current mainstay of tuberculosis diagnosis worldwide, detects only half of tuberculosis cases, mycobacterial culture can take weeks to provide a diagnosis, and rapid techniques such as nucleic acid amplification require infrastructure that is often not available in developing countries. Consequently, in India and other developing countries, serological tests are widely used for the diagnosis of tuberculosis. Serological tests detect antibodies against M. tuberculosis in the blood (antibodies are proteins made by the immune system in response to infections). Serological tests are fast and simple to perform, but they are not recommended for clinical use, and the available evidence suggests that they do not diagnose tuberculosis accurately. Even so, and in the absence of information about the cost and impact (cost-effectiveness) of serological testing, about 1.5 million serological tests for tuberculosis are conducted every year in India at a cost of more than US$15 million. Here, the researchers analyze the cost-effectiveness of serological tests compared to other diagnostic tests from the perspective of tuberculosis control in India.
What Did the Researchers Do and Find?
The researchers used "decision analysis" to estimate the cost-effectiveness of sputum smear microscopy, microscopy plus liquid culture using the MGIT system, and serological testing using the widely used anda-tb ELISA commercial test in a hypothetical group of 1.5 million people suspected of having tuberculosis. Decision analysis formally assesses the decision-making process by using models that evaluate outcomes under different scenarios. By feeding data on the costs and accuracy of different diagnostic tests into their decision-analysis model, the researchers estimate that, over a year, serology would generate 14,000 more tuberculosis diagnoses than sputum microscopy. However, it would also generate 121,000 more false-positive diagnoses and 32,000 more tuberculosis transmissions to other people (secondary transmissions), and avert 102,000 fewer disability-adjusted life years (DALYs; a DALY is a year of healthy life lost because of premature death or disability) at four times the incremental cost of sputum microscopy. MGIT culture added to sputum smear microscopy would avert 130,000 DALYs at an incremental cost of US$213 per DALY averted. Finally, sensitivity analyses (reruns of the decision-analysis model using different values for test costs and accuracy) identified no scenario in which serology was either less costly or more effective than sputum smear microscopy alone or in which serology plus sputum microscopy was more cost-effective than MGIT culture plus sputum microscopy.
What Do These Findings Mean?
These findings identify sputum smear microscopy as the most cost-effective existing diagnostic test for tuberculosis in India. Moreover, they suggest that in areas where high-quality microscopy is available, resources are sufficient, and infrastructure to effectively use culture exists, the addition of MGIT culture to sputum smear microscopy would be more cost-effective than the addition of serology. Importantly, these findings suggest that, if used as an initial test for tuberculosis in India, serology would result in more DALYs, more secondary infections, and more false-positive diagnoses than sputum smear microscopy while increasing per-patient costs to the Indian tuberculosis control sector. Given these findings and the results of a recent updated systematic review on the accuracy of serological tests, the World Health Organization's Strategic and Technical Advisory Group for Tuberculosis recently advised against the use of currently available serological tests for the diagnosis of tuberculosis. The WHO negative policy against serological tests must now be implemented in India.
Additional Information
Please access these Web sites via the online version of this summary at http://dx.doi.org/10.1371/journal.pmed.1001074.
Details of the recent systematic review of serological tests for tuberculosis diagnosis are available in a PLoS Medicine Research Article by Steingart et al.
The World Health Organization provides information on all aspects of tuberculosis, including tuberculosis diagnostics and the Stop TB Partnership (some information is in several languages); its Strategic and Technical Advisory Group for Tuberculosis recommendations on tuberculosis diagnosis are available
The Evidence-based TB Diagnosis Web site by the Stop TB Partnership's New Diagnostics Working Group provides evidence syntheses on various TB tests, along with guidelines, resources, and training materials
The US Centers for Disease Control and Prevention has information about tuberculosis, including information on the diagnosis of tuberculosis disease
The US National Institute of Allergy and Infectious Diseases also has information on all aspects of tuberculosis
MedlinePlus has links to further information about tuberculosis (in English and Spanish)
Related Articles
Electrophoretic Analysis of Indian Isolates of Mycoplasma agalactiae and Mycoplasma bovis by SDS-PAGE and Immunoblotting
Kumar, Amit
Srivastava, N. C.
Singh, V. P.
Sunder, Jai
Veterinary Medicine International
Mycoplasma agalactiae and Mycoplasma bovis both are responsible for respiratory conditions in sheep and goats. M. agalactiae is a major pathogen of sheep and goats and accounts for almost 90% of outbreaks of contagious agalactia syndrome in goats and almost 100% in sheep. On the basis of clinical signs and cultural, morphological, and biochemical characterization it is almost impossible to differentiate between both the species. Moreover, due to presence of genomic and proteomic similarity most of the time routine diagnostic tests fail to differentiate between them. Hence the present study was conducted to find out the protein profile of isolates of both the species by SDS-PAGE and to find out the cross-reacting as well as differentiating immunogenic proteins by Immunoblotting, which can be of immunoprophylactic as well as diagnostic values. The study revealed 6-7 major immunogenic cross-reactive proteins with the presence of two important non-cross-reacting species specific polypeptides particularly 25.50 and 24.54 kDa in M. agalactiae and M. bovis, respectively, that might be of diagnostic values.
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Sensitive and Specific Enzyme-Linked Immunosorbent Assay for Detecting Serum Antibodies against Mycobacterium avium subsp. paratuberculosis in Fallow Deer
Prieto, José M.
Balseiro, Ana
Casais, Rosa
Abendaño, Naiara
Fitzgerald, Liam E.
Garrido, Joseba M.
Juste, Ramon A.
Alonso-Hearn, Marta
Clinical and Vaccine Immunology : CVI
The enzyme-linked immunosorbent assay (ELISA) is the diagnostic test most commonly used in efforts to control paratuberculosis in domestic ruminants. However, commercial ELISAs have not been validated for detecting antibodies against Mycobacterium avium subsp. paratuberculosis in wild animals. In this study, we compared the sensitivities and specificities of five ELISAs using individual serum samples collected from 41 fallow deer with or without histopathological lesions consistent with paratuberculosis. Two target antigenic preparations were selected, an ethanol-treated protoplasmic preparation obtained from a fallow deer M. avium subsp. paratuberculosis isolate (ELISAs A and B) and a paratuberculosis protoplasmic antigen (PPA3) (ELISAs C and D). Fallow deer antibodies bound to the immobilized antigens were detected by using a horseradish peroxidase (HRP)-conjugated anti-fallow deer IgG antibody (ELISAs A and C) or HRP-conjugated protein G (ELISAs B and D). A commercially available assay, ELISA-E, which was designed to detect M. avium subsp. paratuberculosis antibodies in cattle, sheep, and goats, was also tested. Although ELISAs A, C, and E had the same sensitivity (72%), ELISAs A and C were more specific (100%) for detecting fallow deer with lesions consistent with paratuberculosis at necropsy than was the ELISA-E (87.5%). In addition, the ELISA-A was particularly sensitive for detecting fallow deer in the latent stages of infection (62.5%). The antibody responses detected with the ELISA-A correlated with both the severity of enteric lesions and the presence of acid-fast bacteria in gut tissue samples. In summary, our study shows that the ELISA-A can be a cost-effective diagnostic tool for preventing the spread of paratuberculosis among fallow deer populations.
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An international collaborative study to determine the prevalence of contagious caprine pleuropneumonia by monoclonal antibody-based cELISA
Peyraud, Armelle
Poumarat, François
Tardy, Florence
Manso-Silván, Lucía
Hamroev, Karomatullo
Tilloev, Tillo
Amirbekov, Mullojon
Tounkara, Karim
Bodjo, Charles
Wesonga, Hezron
Nkando, Isabel Gacheri
Jenberie, Shiferaw
Yami, Martha
Cardinale, Eric
Meenowa, Deodass
Jaumally, Mahmad Reshad
Yaqub, Tahir
Shabbir, Muhammad Zubair
Mukhtar, Nadia
Halimi, Mohibullah
Ziay, Ghulam Mohammad
Schauwers, Willy
Noori, Hafizullah
Rajabi, Ali Madad
Ostrowski, Stéphane
Thiaucourt, François
BMC Veterinary Research
Few serological tests are available for detecting antibodies against Mycoplasma capricolum subsp. capripneumoniae, the causal agent of contagious caprine pleuropneumonia (CCPP). The complement fixation test, the test prescribed for international trade purposes, uses a crude antigen that cross-reacts with all the other mycoplasma species of the "mycoides cluster" frequently infecting goat herds. The lack of a more specific test has been a real obstacle to the evaluation of the prevalence and economic impact of CCPP worldwide. A new competitive ELISA kit for CCPP, based on a previous blocking ELISA, was formatted at CIRAD and used to evaluate the prevalence of CCPP in some regions of Kenya, Ethiopia, Mauritius, Tajikistan and Pakistan in an international collaborative study.
The strict specificity of the test was confirmed in CCPP-free goat herds exposed to other mycoplasma species of the "mycoides cluster". Prevalence studies were performed across the enzootic range of the disease in Africa and Asia. Seroprevalence was estimated at 14.6% in the Afar region of Ethiopia, whereas all the herds presented for CCPP vaccination in Kenya tested positive (individual seroprevalence varied from 6 to 90% within each herd). In Mauritius, where CCPP emerged in 2009, nine of 62 herds tested positive. In Central Asia, where the disease was confirmed only recently, no positive animals were detected in the Wakhan District of Afghanistan or across the border in neighboring areas of Tajikistan, whereas seroprevalence varied between 2.7% and 44.2% in the other districts investigated and in northern Pakistan. The test was also used to monitor seroconversion in vaccinated animals.
This newly formatted CCPP cELISA kit has retained the high specificity of the original kit. It can therefore be used to evaluate the prevalence of CCPP in countries or regions without vaccination programs. It could also be used to monitor the efficacy of vaccination campaigns as high-quality vaccines induce high rates of seroconversion.
Contagious caprine pleuropneumonia; Competitive ELISA; Seroprevalence; Kenya; Ethiopia; Mauritius; Tajikistan; Afghanistan; Pakistan; Vaccine quality control
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Commercial Serological Tests for the Diagnosis of Active Pulmonary and Extrapulmonary Tuberculosis: An Updated Systematic Review and Meta-Analysis
Steingart, Karen R.
Flores, Laura L.
Dendukuri, Nandini
Schiller, Ian
Laal, Suman
Ramsay, Andrew
Hopewell, Philip C.
Pai, Madhukar
PLoS Medicine
An up-to-date systematic review and meta-analysis by Karen Steingart and colleagues confirms that commercially available serological tests do not provide an accurate diagnosis of tuberculosis.
Serological (antibody detection) tests for tuberculosis (TB) are widely used in developing countries. As part of a World Health Organization policy process, we performed an updated systematic review to assess the diagnostic accuracy of commercial serological tests for pulmonary and extrapulmonary TB with a focus on the relevance of these tests in low- and middle-income countries.
Methods and Findings
We used methods recommended by the Cochrane Collaboration and GRADE approach for rating quality of evidence. In a previous review, we searched multiple databases for papers published from 1 January 1990 to 30 May 2006, and in this update, we add additional papers published from that period until 29 June 2010. We prespecified subgroups to address heterogeneity and summarized test performance using bivariate random effects meta-analysis. For pulmonary TB, we included 67 studies (48% from low- and middle-income countries) with 5,147 participants. For all tests, estimates were variable for sensitivity (0% to 100%) and specificity (31% to 100%). For anda-TB IgG, the only test with enough studies for meta-analysis, pooled sensitivity was 76% (95% CI 63%–87%) in smear-positive (seven studies) and 59% (95% CI 10%–96%) in smear-negative (four studies) patients; pooled specificities were 92% (95% CI 74%–98%) and 91% (95% CI 79%–96%), respectively. Compared with ELISA (pooled sensitivity 60% [95% CI 6%–65%]; pooled specificity 98% [95% CI 96%–99%]), immunochromatographic tests yielded lower pooled sensitivity (53%, 95% CI 42%–64%) and comparable pooled specificity (98%, 95% CI 94%–99%). For extrapulmonary TB, we included 25 studies (40% from low- and middle-income countries) with 1,809 participants. For all tests, estimates were variable for sensitivity (0% to 100%) and specificity (59% to 100%). Overall, quality of evidence was graded very low for studies of pulmonary and extrapulmonary TB.
Despite expansion of the literature since 2006, commercial serological tests continue to produce inconsistent and imprecise estimates of sensitivity and specificity. Quality of evidence remains very low. These data informed a recently published World Health Organization policy statement against serological tests.
Please see later in the article for the Editors' Summary
Editors' Summary
Every year nearly 10 million people develop tuberculosis—a contagious bacterial infection—and about two million people die from the disease. Mycobacterium tuberculosis, the bacterium that causes tuberculosis, is spread in airborne droplets when people with the disease cough or sneeze. It usually infects the lungs (pulmonary tuberculosis) but can also infect the lymph nodes, bones, and other tissues (extrapulmonary tuberculosis). The characteristic symptoms of tuberculosis are a persistent cough, weight loss, and night sweats. Diagnostic tests for the disease include microscopic examination of sputum (mucus brought up from the lungs by coughing) for M. tuberculosis bacilli, chest radiography, mycobacterial culture (in which bacteriologists try to grow M. tuberculosis from sputum or tissue samples), and nucleic acid amplification tests (which detect the bacterium's genome in patient samples). Tuberculosis can usually be cured by taking several powerful drugs daily or several times a week for at least six months.
Why Was This Study Done?
Although efforts to control tuberculosis have advanced over the past decade, missed tuberculosis diagnoses and mismanaged tuberculosis continue to fuel the global epidemic. A missed diagnosis may lead to more severe illness and death, especially for people infected with both tuberculosis and HIV. Also, a missed diagnosis means that an untreated individual with pulmonary tuberculosis may remain infectious for longer, continuing to spread tuberculosis within the community Missed diagnoses are a particular problem in resource-limited countries where sputum microscopy and chest radiography often perform poorly and other diagnostic tests are too expensive and complex for routine use. Serological tests, which detect antibodies against M. tuberculosis in the blood (antibodies are proteins made by the immune system in response to infections), might provide a way to diagnose tuberculosis in resource-limited countries. Indeed, many serological tests for tuberculosis diagnosis are on sale in developing countries. However, because of doubts about the accuracy of these commercial tests, they are not recommended for use in routine practice. In this systematic review and meta-analysis, the researchers assess the diagnostic accuracy of commercial serological tests for pulmonary and extrapulmonary tuberculosis. A systematic review uses predefined criteria to identify all the research on a given topic; meta-analysis is a statistical method that combines the results of several studies.
What Did the Researchers Do and Find?
The researchers searched the literature for studies that evaluated serological tests for active tuberculosis published between 1990 and 2010. They used data from these studies to calculate each test's sensitivity (the proportion of patients with a positive serological test among patients with tuberculosis confirmed by a reference method; a high sensitivity indicates that the test detects most patients with tuberculosis) and specificity (the proportion of patients with a negative serological result among people without tuberculosis; a high specificity means the test gives few false-positive diagnoses). They also assessed the methodological quality of each study and rated the overall quality of the evidence. The researchers found 67 studies (half from low/middle-income countries) that evaluated serological tests for the diagnosis of pulmonary tuberculosis. The sensitivity of these tests varied between studies, ranging from 0% to 100%; their specificities ranged from 31% to 100%. For the anda-TB IgG test—the only test with sufficient studies for a meta-analysis—the pooled sensitivity from the relevant studies was 76% in smear-positive patients and 59% in smear-negative patients. The pooled specificities were 92% and 91%, respectively. The researchers found 25 studies (40% from low/middle-income countries) that evaluated serological tests for the diagnosis of extrapulmonary tuberculosis. Again, sensitivities and specificities for each test varied greatly between studies, ranging from 0% to 100% and 59% to 100%, respectively. Overall, for both pulmonary and extrapulmonary tuberculosis, the quality of evidence from the studies of the serological tests was graded very low.
What Do These Findings Mean?
This systematic review, which updates an analysis published in 2007, indicates that commercial serological tests do not provide an accurate diagnosis of tuberculosis. This finding confirms previous systematic reviews of the evidence, despite a recent expansion in the relevant literature. Moreover, the researchers' analysis indicates that the overall quality of the body of evidence on these tests remains poor. Many of the identified studies used unsatisfactory patient selection methods, for example. Clearly, there is a need for continued and improved research on existing serological tests and for research into new approaches to the serological diagnosis of tuberculosis. For now, though, based on these findings, cost-effectiveness data, and expert opinion, the World Health Organization has issued a recommendation against the use of currently available serological tests for the diagnosis of tuberculosis, while stressing the importance of continued research on these and other tests that could provide quick and accurate diagnosis of TB.
Additional Information
Please access these Web sites via the online version of this summary at http://dx.doi.org/10.1371/journal.pmed.1001062.
The World Health Organization provides information on all aspects of tuberculosis, including information on tuberculosis diagnostics on the Stop TB Partnership (some information is in several languages); the Strategic and Technical Advisory Group for Tuberculosis recommendations on tuberculosis diagnosis are available
The Web site Evidence-Based Tuberculosis Diagnosis (from Stop TB Partnership's New Diagnostics Working Group) provides access to several resources on TB diagnostics, including systematic reviews, guidelines, and training materials
The US Centers for Disease Control and Prevention has information about tuberculosis, including information on the diagnosis of tuberculosis disease
The US National Institute of Allergy and Infectious Diseases also has detailed information on all aspects of tuberculosis
MedlinePlus has links to further information about tuberculosis (in English and Spanish)
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Evaluation of Two New Commercial Tests for the Diagnosis of Acute Dengue Virus Infection Using NS1 Antigen Detection in Human Serum
Dussart, Philippe
Petit, Laure
Labeau, Bhety
Bremand, Laetitia
Leduc, Alexandre
Moua, David
Matheus, Séverine
Baril, Laurence
PLoS Neglected Tropical Diseases
We compared the performance of two new commercial tests for the detection of dengue NS1 protein during the clinical phase of dengue virus (DENV) infection—an immunochromatographic test allowing rapid detection of the NS1 antigen, Dengue NS1 Ag STRIP (Bio-Rad Laboratories - Marnes La Coquette, France), and a two-step sandwich-format microplate enzyme-linked immunosorbent assay (ELISA), pan-E Dengue Early ELISA (Panbio - Brisbane, Australia)—with a one-step sandwich-format microplate ELISA, the Platelia Dengue NS1 Ag test (Bio-Rad).
We tested 272 serum samples from patients with dengue disease. Of these, 222 were from patients with acute infection of one of the four dengue serotypes, detected by RT-PCR and/or virus isolation. Forty-eight acute-phase serum samples from patients not infected with dengue virus were also included.
The sensitivity of the Platelia Dengue NS1 Ag test on acute serum samples (n = 222) was 87.4% (95% confidence interval: 82.3% to 91.5%); that of Dengue NS1 Ag STRIP was 81.5% (95% CI: 75.8% to 86.4%) after 15 minutes and 82.4% (95% CI: 76.8% to 87.2%) after 30 minutes. Both tests had a specificity of 100% (97.5% CI, one-sided test: 92.6% to 100.0%). The pan-E Dengue Early ELISA had a sensitivity of 60.4% (95% CI: 53.4% to 66.8%) and a specificity of 97.9% (95% CI: 88.9% to 99.9%).
Our findings support the use of diagnostic tools based on the NS1 antigen detection for the diagnosis of acute DENV infection. The immunochromatographic test, Dengue NS1 Ag STRIP—the first rapid diagnostic test for DENV infection—was highly sensitive and specific, and would therefore be a suitable first-line test in the field. The pan-E Dengue Early ELISA was less sensitive than the Platelia test; this two-step ELISA should be combined with DENV IgM antibody detection for the diagnosis of DENV infection.
Author Summary
Dengue is a viral disease transmitted by mosquitoes that is endemic in more than 100 countries in tropical areas, threatening over 2.5 billion people. It causes a wide range of symptoms and has severe forms. In reference laboratories, dengue disease is confirmed by virus isolation or genome detection during the acute phase, and by serological methods during the early convalescent phase. The viral NS1 protein circulates in the sera of infected patients throughout the clinical phase of the disease. Novel diagnostic tests based on NS1 detection have been recently developed and marketed. We compared the performance of two tests for detecting dengue NS1 protein during the clinical phase of dengue infection (an immunochromatographic test (ICT) from Bio-Rad allowing rapid detection of the NS1 antigen and a two-step sandwich-format ELISA from Panbio) with the one-step sandwich-format microplate ELISA (Bio-Rad). The ICT test performed better than the ELISA test from Panbio. This study confirms that diagnostic tests based on NS1 could be used in routine clinical practice in poorly equipped laboratories and that dengue diagnosis could therefore be confirmed without the need for testing in reference laboratories. This represents a crucial step towards the control of dengue disease in the human population.
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Evaluation of a commercial ELISA kit for detection of antibodies against Toxoplasma gondii in serum, plasma and meat juice from experimentally and naturally infected sheep
Glor, Sabine B
Edelhofer, Renate
Grimm, Felix
Deplazes, Peter
Basso, Walter
Parasites & Vectors
Toxoplasmosis is one of the most common food borne zoonoses worldwide, and can be a serious life-threatening disease in the congenitally infected fetus and in immunosupressed patients. Among food animals, sheep along with goats and pigs possess the highest incidence of T. gondii cysts in meat, and play a major role as a source of human infection.
In this study, a new commercial ELISA kit (PrioCHECK® Toxoplasma Ab SR, Prionics Schlieren-Zurich, Switzerland) for the detection of anti-T. gondii antibodies in serum, plasma and meat juice of sheep, was evaluated by comparing it with the indirect fluorescent antibody test (IFAT), indirect haemagglutination test (IHA) and real-time PCR, on samples from experimentally inoculated and naturally exposed sheep.
The commercial ELISA detected the infection status in 50% and 100% of sheep orally inoculated with 10,000 T. gondii oocysts (n = 6), from two or three weeks post infection (wpi), respectively, both on serum and plasma samples. Meat juice from all experimentally inoculated sheep collected at slaughter (12 wpi) showed positive ELISA values. In naturally exposed sheep (n = 396), the ELISA showed a very good agreement with IFAT (kappa = 0.91-1.0) and IHA (kappa = 0.96-1.0) performed on serum; and a positive correlation was observed between ELISA values and IFAT titers. By a Receiver Operating Characteristics (ROC) curve analysis, the commercial ELISA had relative sensitivities between 93.33% and 100%, and relative specificities between 96.87% and 100% respect to IFAT and IHA, depending on the considered cut-off value and animal groups tested. Furthermore, the ELISA correctly recognized all animals reacting positive in real-time PCR. The ELISA results on meat juice agreed with those on serum samples in all experimentally inoculated animals, and in 94 out of 96 (97.9%) naturally exposed sheep, when meat juice was tested at a 1:10 dilution.
The commercial ELISA kit evaluated in this study could represent a valuable tool to improve the surveillance and reporting system for T. gondii in sheep populations at the farm level or for diagnosis at the slaughterhouse, contributing to the control of this widespread zoonosis.
Toxoplasma gondii; Sheep; Diagnosis; ELISA; IFAT; Indirect haemagglutination; Antibodies; Meat juice
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Use of Recombinant Antigens To Detect Antibodies against Mycoplasma suis, with Correlation of Serological Results to Hematological Findings▿
Hoelzle, Katharina
Grimm, Julia
Ritzmann, Mathias
Heinritzi, Karl
Torgerson, Paul
Hamburger, Anja
Wittenbrink, Max M.
Hoelzle, Ludwig E.
Clinical and Vaccine Immunology : CVI
Porcine eperythrozoonosis is a disease with worldwide distribution caused by the unculturable hemotrophic bacterium Mycoplasma suis. Current serological testing utilizes crude M. suis antigens purified from the blood of experimentally infected pigs. These antigens show high variability and are restricted to specialized laboratories. We evaluated a novel serological assay based on two recombinant M. suis antigens (rMSG1 and rHspA1). Antigen specificity was proven by means of sera raised against nonhemotrophic mycoplasma and other relevant bacteria. Using experimental and convalescent-phase sera, rMSG1 and rHspA1 enzyme-linked immunosorbent assays (ELISAs) demonstrated sensitivities, specificities, and predictive values (94.0 to 100.0%) equal to or higher than those of the M. suis whole-cell ELISA. Field samples from 120 weaning piglets grouped by quantitative PCR results were used to evaluate the diagnostic capability of the new ELISA systems in comparison to that of the whole-cell ELISA. Assuming a 100.0% specificity of the PCR, the whole-cell ELISA, rHspA1 ELISA, and rMSG1 ELISA showed specificities of 84.8%, 83.8%, and 90.6% and sensitivities of 61.5%, 74.0% and 58.1%, respectively. Cohen's kappa coefficients comparing the recombinant ELISAs to the whole-cell ELISA indicate moderate to substantial agreement. The detection of anti-MSG1 and/or anti-HspA1 antibodies in pigs was significantly correlated with decreased hematocrit, erythrocyte numbers, and hemoglobin concentrations, indicating that a single seropositive result is connected with clinical and etiological significance. In conclusion, rMSG1 and rHspA1 are sensitive and specific serological and infection markers which are for the first time used independently of animal experiments. They are especially fit to be used in routine diagnosis, pathogenesis studies, and large-scale epidemiological investigations.
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Mapping Antigenic Sites of an Immunodominant Surface Lipoprotein of Mycoplasma agalactiae, AvgC, with the Use of Synthetic Peptides
Santona, Antonella
Carta, Franco
Fraghí, Peppinetta
Turrini, Franco
Infection and Immunity
As a first step toward the design of an epitope vaccine to prevent contagious agalactia, the strongly immunogenic 55-kDa protein of Mycoplasma agalactiae was studied and found to correspond to the AvgC protein encoded by the avgC gene. The avg genes of M. agalactiae, which encode four variable surface lipoproteins, display a significant homology to the vsp (variable membrane surface lipoproteins) genes of the bovine pathogen Mycoplasma bovis at their promoter region as well as their N-terminus-encoding regions. Some members of the Vsp family are known to be involved in cytoadhesion to host cells. In order to localize immunogenic peptides in the AvgC antigen, the protein sequence was submitted to epitope prediction analysis, and five sets of overlapping peptides, corresponding to five selected regions, were synthesized by Spot synthesis. Reactive peptides were selected by immunobinding assay with sera from infected sheep. The three most immunogenic epitopes were shown to be surface exposed by immunoprecipitation assays, and one of these was specifically recognized by all tested sera. Our study indicates that selected epitopes of the AvgC lipoprotein may be used to develop a peptide-based vaccine which is effective against M. agalactiae infection.
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The effect of pre-enrichment on recovery of Streptococcus agalactiae, Staphylococcus aureus and mycoplasma from bovine milk.
Thurmond, M. C.
Tyler, J. W.
Luiz, D. M.
Holmberg, C. A.
Picanso, J. P.
Epidemiology and Infection
The study was conducted to determine whether pre-enrichment would increase sensitivity of detecting Streptococcus (Str.) agalactiae, Staphylococcus (S.) aureus, and mycoplasma in bovine milk. Two procedures were followed, one involving direct inoculation of milk on bovine blood agar, and the other involving preenrichment in broth followed by inoculation on agar. Logistic regression was used to predict the probability of isolation as a function of culture procedure and two additional covariates, the California Mastitis Test (CMT) score of the milk and the type of sample (indicating sample storage temperature and herd mastitis status). A total of 13778 milk samples was cultured for each of the three bacteria. By using results of both direct inoculation and pre-enrichment, the probability of isolation compared to use of direct inoculation only and adjusted for effects of other variables was increased 3.6-fold for Str. agalactiae, 1.6-fold for S. aureus and 1.7-fold for mycoplasma. The probability of isolation for all three bacteria increased as the CMT score increased. For Str. agalactiae, there was a statistical interaction predicting that enrichment improved the odds of isolation more from milk with high CMT scores than from milk with low scores. Results indicate that pre-enrichment can substantially increase the sensitivity of bacteriological screening of dairy cows for mastitis caused by Str. agalactiae, S. aureus, and mycoplasma.
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Emergence of Atypical Mycoplasma agalactiae Strains Harboring a New Prophage and Associated with an Alpine Wild Ungulate Mortality Episode
Tardy, Florence
Baranowski, Eric
Nouvel, Laurent-Xavier
Mick, Virginie
Manso-Silvàn, Lucía
Thiaucourt, François
Thébault, Patricia
Breton, Marc
Sirand-Pugnet, Pascal
Blanchard, Alain
Garnier, Alexandre
Gibert, Philippe
Game, Yvette
Poumarat, François
Citti, Christine
Applied and Environmental Microbiology
The bacterium Mycoplasma agalactiae is responsible for contagious agalactia (CA) in small domestic ruminants, a syndrome listed by the World Organization for Animal Health and responsible for severe damage to the dairy industry. Recently, we frequently isolated this pathogen from lung lesions of ibexes during a mortality episode in the French Alps. This situation was unusual in terms of host specificity and tissue tropism, raising the question of M. agalactiae emergence in wildlife. To address this issue, the ibex isolates were characterized using a combination of approaches that included antigenic profiles, molecular typing, optical mapping, and whole-genome sequencing. Genome analyses showed the presence of a new, large prophage containing 35 coding sequences (CDS) that was detected in most but not all ibex strains and has a homolog in Mycoplasma conjunctivae, a species causing keratoconjunctivitis in wild ungulates. This and the presence in all strains of large integrated conjugative elements suggested highly dynamic genomes. Nevertheless, M. agalactiae strains circulating in the ibex population were shown to be highly related, most likely originating from a single parental clone that has also spread to another wild ungulate species of the same geographical area, the chamois. These strains clearly differ from strains described in Europe so far, including those found nearby, before CA eradication a few years ago. While M. agalactiae pathogenicity in ibexes remains unclear, our data showed the emergence of atypical strains in Alpine wild ungulates, raising the question of a role for the wild fauna as a potential reservoir of pathogenic mycoplasmas.
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The use of a Psoroptes ovis serodiagnostic test for the analysis of a natural outbreak of sheep scab
Burgess, Stewart TG
Innocent, Giles
Nunn, Francesca
Frew, David
Kenyon, Fiona
Nisbet, Alasdair J
Huntley, John F
Parasites & Vectors
Sheep scab is a highly contagious disease of sheep caused by the ectoparasitic mite Psoroptes ovis. The disease is endemic in the UK and has significant economic impact through its effects on performance and welfare. Diagnosis of sheep scab is achieved through observation of clinical signs e.g. itching, pruritis and wool loss and ultimately through the detection of mites in skin scrapings. Early stages of infestation are often difficult to diagnose and sub-clinical animals can be a major factor in disease spread. The development of a diagnostic assay would enable farmers and veterinarians to detect disease at an early stage, reducing the risk of developing clinical disease and limiting spread.
Serum samples were obtained from an outbreak of sheep scab within an experimental flock (n = 480 (3 samples each from 160 sheep)) allowing the assessment, by ELISA of sheep scab specific antibody prior to infestation, mid-outbreak (combined with clinical assessment) and post-treatment.
Analysis of pre-infestation samples demonstrated low levels of potential false positives (3.8%). Of the 27 animals with clinical or behavioural signs of disease 25 tested positive at the mid-outbreak sampling period, however, the remaining 2 sheep tested positive at the subsequent sampling period. Clinical assessment revealed the absence of clinical or behavioural signs of disease in 132 sheep, whilst analysis of mid-outbreak samples showed that 105 of these clinically negative animals were serologically positive, representing potential sub-clinical infestations.
This study demonstrates that this ELISA test can effectively diagnose sheep scab in a natural outbreak of disease, and more importantly, highlights its ability to detect sub-clinically infested animals. This ELISA, employing a single recombinant antigen, represents a major step forward in the diagnosis of sheep scab and may prove to be critical in any future control program.
Ectoparasite; Diagnostic; ELISA; Psoroptes ovis; Sheep scab
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Gupta, Shivali
Wan, Xianxiu
Zago, Maria P.
Martinez Sellers, Valena C.
Silva, Trevor S.
Assiah, Dadjah
Dhiman, Monisha
Nuñez, Sonia
Petersen, John R.
Vázquez-Chagoyán, Juan C.
Estrada-Franco, Jose G.
Garg, Nisha Jain
PLoS Neglected Tropical Diseases
Chagas disease, caused by Trypanosoma cruzi, is endemic in Latin America and an emerging infectious disease in the US and Europe. We have shown TcG1, TcG2, and TcG4 antigens elicit protective immunity to T. cruzi in mice and dogs. Herein, we investigated antigenicity of the recombinant proteins in humans to determine their potential utility for the development of next generation diagnostics for screening of T. cruzi infection and Chagas disease.
Methods and Results
Sera samples from inhabitants of the endemic areas of Argentina-Bolivia and Mexico-Guatemala were analyzed in 1st-phase for anti-T. cruzi antibody response by traditional serology tests; and in 2nd-phase for antibody response to the recombinant antigens (individually or mixed) by an ELISA. We noted similar antibody response to candidate antigens in sera samples from inhabitants of Argentina and Mexico (n = 175). The IgG antibodies to TcG1, TcG2, and TcG4 (individually) and TcGmix were present in 62–71%, 65–78% and 72–82%, and 89–93% of the subjects, respectively, identified to be seropositive by traditional serology. Recombinant TcG1- (93.6%), TcG2- (96%), TcG4- (94.6%) and TcGmix- (98%) based ELISA exhibited significantly higher specificity compared to that noted for T. cruzi trypomastigote-based ELISA (77.8%) in diagnosing T. cruzi-infection and avoiding cross-reactivity to Leishmania spp. No significant correlation was noted in the sera levels of antibody response and clinical severity of Chagas disease in seropositive subjects.
Three candidate antigens were recognized by antibody response in chagasic patients from two distinct study sites and expressed in diverse strains of the circulating parasites. A multiplex ELISA detecting antibody response to three antigens was highly sensitive and specific in diagnosing T. cruzi infection in humans, suggesting that a diagnostic kit based on TcG1, TcG2 and TcG4 recombinant proteins will be useful in diverse situations.
Author Summary
Chagas disease is the most common cause of congestive heart failure related deaths among young adults in the endemic areas of South and Central America and Mexico. Diagnosis and treatment of T. cruzi infection has remained difficult and challenging after 100 years of its identification. In >95% of human cases, T. cruzi infection remains undiagnosed until several years later when chronic evolution of progressive disease results in clinical symptoms associated with cardiac damage. Diagnosis generally depends on the measurement of T. cruzi–specific antibodies that can result in false positives. A conclusive diagnosis of T. cruzi infection thus often requires multiple serological tests, in combination with epidemiological data and clinical symptoms. In this study, we investigated the antibody response to TcG1, TcG2, and TcG4 in clinically characterized chagasic patients. These antigens were identified as vaccine candidates and shown to elicit protective immunity to T. cruzi and Chagas disease in experimental animals. Our data show the serology test developed using the TcGmix (multiplex ELISA) is a significantly better alternative to epimastigote extracts currently used in T. cruzi serodiagnosis or the trypomastigote lysate used in this study for comparison purposes.
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Serological testing of cattle experimentally infected with Mycoplasma mycoides subsp. mycoides Small Colony using four different tests reveals a variety of seroconversion patterns
Schubert, Evelyn
Sachse, Konrad
Jores, Jörg
Heller, Martin
BMC Veterinary Research
To study the specific antibody response to infection with Mycoplasma mycoides subsp. mycoides Small Colony (MmmSC), the agent of Contagious Bovine Pleuropneumonia (CBPP), we examined three panels of sera collected during three experimental infection trials in African cattle. The methods used included an in-house complement fixation test (CFT), a commercially available CFT, a competitive antibody ELISA (cELISA) and the immunoblotting test (IBT). In addition, lung tissue samples were examined by culture.
A total of 89% (51/59) of all experimentally infected animals tested positive on at least one of the serological tests throughout the trial. The specific antibody titres to the MmmSC infection became positive first by CFT (6 to 9 days post infection [dpi]), followed by IBT (9 to 13 dpi) and cELISA (13 to 16 dpi). Individual animals were found to display remarkably distinct seroconversion patterns, which allowed their classification into i) early high responders, ii) late high responders, and iii) low responders. In accordance with other studies, none of the present serological tests was capable of detecting all CBPP infected animals.
Comparison of the assays' performance in terms of sensitivity and specificity raises serious questions as to their reliability for identification of infected individuals in the field. In view of these limitations, a combination of CFT and cELISA can markedly improve CBPP diagnosis at single-animal level.
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Surveys on Coxiella burnetii infections in Swedish cattle, sheep, goats and moose
Ohlson, Anna
Malmsten, Jonas
Frössling, Jenny
Bölske, Göran
Aspán, Anna
Dalin, Anne-Marie
Lindberg, Ann
Acta Veterinaria Scandinavica
Q fever is a zoonotic disease caused by the bacterium Coxiella burnetii. Prevalence data in ruminant species are important to support risk assessments regarding public and animal health. The aim was to investigate the presence of or exposure to C. burnetii in cattle, sheep, goats and moose, and to compare two enzyme-linked immunosorbent assays (ELISAs). National surveys of antibodies against C. burnetii were performed for dairy cattle (n=1537), dairy goats (n=58) and sheep (n=518). Bovine samples consisted of bulk milk, caprine of pooled milk, and ovine of pooled serum. Antibodies were investigated in moose samples (n=99) from three regions. A one-year regional cattle bulk milk survey was performed on the Isle of Gotland (n=119, four occasions). Cattle, sheep and goat samples were analysed with indirect ELISA and moose samples with complement fixation test. For the sheep, goat, and parts of the cattle survey, samples were run in parallel by ELISAs based on antigens from infected ruminants and ticks. Bulk milk samples from the regional cattle survey and vaginal swabs from a subset of the sheep herds (n=80) were analysed for the agent by polymerase chain reaction. Spatial clustering was investigated in the national cattle survey.
The prevalence of antibodies in dairy herds was 8.2% with large regional differences. High risk clusters were identified in the southern regions. The prevalence among dairy herds on the Isle of Gotland varied from 55.9% to 64.6% and 46.4% to 58.9.0% for antibodies and agent, respectively, overall agreement between agent and antibodies was 85.2%. The prevalence of antibodies in sheep was 0.6%, the agent was not detected the vaginal swabs. Antibodies were not detected in goats or moose, although parts of the moose samples were collected in an area with high prevalence in cattle. The overall agreement between the two ELISAs was 90.4%.
The prevalence of antibodies against C. burnetii in dairy cattle in Sweden shows large regional differences. The results suggest that C. burnetii is a rare pathogen among Swedish moose, dairy goat and sheep. ELISAs based on ruminant and tick antigen performed in a similar manner under Swedish conditions.
Coxiella burnetii; Surveillance; Epidemiology; Cattle; Goat; Sheep; Moose; Cervids; Antibodies; Test evaluation
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Mycoplasma agalactiae, an Etiological Agent of Contagious Agalactia in Small Ruminants: A Review
Kumar, Amit
Rahal, Anu
Chakraborty, Sandip
Verma, Amit Kumar
Dhama, Kuldeep
Veterinary Medicine International
Mycoplasma agalactiae is one of the causal agents of classical contagious agalactia (CA), a serious, economically important but neglected enzootic disease of small ruminants. It occurs in many parts of the world and most notably in the Mediterranean Basin. Following the infection common complications are septicaemia, mastitis, arthritis, pleurisy, pneumonia, and keratoconjunctivitis. Primary or tentative diagnosis of the organism is based upon clinical signs. Various serological tests, namely, growth precipitation, immunofluorescence, complement fixation test, haemagglutination inhibition, agglutination, immunodiffusion, enzyme immunoassays, immunoelectrophoresis, blotting techniques, and others, are available. Molecular tools seem to be much more sensitive, specific, and faster and help to differentiate various strains. The real-time PCR, multiplex PCR, quantitative PCR, PCR-RFLP, MLST, and gene probes, complementary to segments of chromosomal DNA or 16S ribosomal RNA (rRNA), have strengthened the diagnosis of M. agalactiae. Both live attenuated and adjuvant (alum precipitated or saponified) inactivated vaccines are available with greater use of inactivated ones due to lack of side effects. The present review discusses the etiology, epidemiology, pathogenesis, and clinical signs of contagious agalactia in small ruminants along with trends and advances in its diagnosis, treatment, vaccination, prevention, and control strategies that will help in countering this disease.
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Science Fiction - A Nerd's Guide to Reading
A Nerd's Guide to Reading
Keywords: Books, Self Improvement, American Lit., Science Fiction
A Nerd's Guide to Reading
Read a great book and wondering what to read next? Ask the famous bookseer
Reading a book is fun, but when you forgot it at home, are bored at work or are just curious to get a sneak peek at whatever you ordered from Amazon, online book sites can be handy too.
Practical Tips for Productive Living
Whether you are just starting out in college or are a more experience learner with years under your belt, there is always more knowledge waiting to be discovered. One great way to do that is to read. This list provides 100 books to expand your knowle...

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- openDemocracy
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About Kuban Taabaldiev
Kuban Taabaldiev was Fulbright Scholar in Journalism in 2007 and now the head of independent news agency ZAMANDASH-PRESS, which focuses on problems of migrants.
Articles by Kuban Taabaldiev
This week's editor
En Liang Khong is openDemocracy's assistant editor.
Digital Liberties
How to fight for digital human rights?
European integration
Six countries. Six different paths to EU integration.


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Hot topics in Business & Enterprise Softwarephp school management system download mlm vb.net school management system school management school fee management system school management software school offline school software for school school report card software student registration system in php css
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Kendall Family History & Genealogy
Find Kendall recordsSearch for Kendall records
Latest Kendall Photos
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Kendall Last Name History & Origin
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Name Origin
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Early Kendalls
These are the earliest records we have of the Kendall family.
Kendall Biographies & Family Trees
Find birth, death records, and obituaries of Kendalls on AncientFaces:
Most Common First Names
- William 3.1%
- John 2.6%
- James 2.5%
- Mary 2.0%
- Robert 1.7%
- George 1.7%
- Thomas 1.6%
- Charles 1.5%
- Edward 1.0%
- Kendall 0.9%
Sample of 16,946 Kendalls bios
Kendall Death Records & Life Expectancy
According to our database of 12,968 people with the last name Kendall that have a birth and death date listed:
Life Expectancy
Oldest Kendalls
These are the longest-lived members of the Kendall family on AncientFaces.
- Ronnie Kendall lived 115 years
- Anna M Kendall lived 105 years
- Jean Kendall lived 105 years
- Ella Kendall lived 104 years
- Garnet Kendall lived 105 years
- Lucille Kendall lived 104 years
- Effie E Kendall lived 105 years
- Clara L Kendall lived 104 years
- Genevieve Kendall lived 103 years
- Cydona E Kendall lived 102 years
Other Kendall Records
Find Kendall recordsSearch for Kendall records
Granny's name was Gertrude Kendall. She also had other sisters and brother(s). At some point Granny married, either in Canada or England ,came to Canada, and gave birth to my mom in 1920-21, Margeurite Gladys Linklater. For some reason unknown to us, Granny remarried a John Aitken from Stratford,Ontario. They lived in Winnipeg, Manitoba, Canada and then moved to Fife Lake, Saskatchewan, Canada and Minto, Manitoba.John Aitken worked with horses in Fife Lake and Minto, farming as well. They had 4 children together, 3 daughters and a son.John (Grandpa) then worked for the Canadian National Railway until his retirement in Winnipeg Manitoba in the 1960's.
A couple of years ago, I was in Manchester and looked up an address I had for a Kendall, but was not successful in getting information that was helpful to my family history search of who my Mother's father was. This Kendall family had owned a cigar shop in Manchester in the past.
I am in England right now visiting and do not have proper info with me that would be useful to add more details. I do have an address and names of people I visited in 2003, and can add this info to my story, if that would be helpful to anyone out there who culd possibly help me. I know this is a far stretch, and very vague, but I am really feeling a need to pursue this.
Please email me at [contact link]
I live in British Columbia, canada and would love to hear formanyone who could help. Thanks a bunch. take care..Betty-Jean
Granny passeda way around 1979 in Winnipeg after a stroke debilitated her.


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FWSA Monday Night Open Mic
Buffalo West Restaurant, 7101 Camp Bowie West, Fort Worth, TX 76116
All songwriters and performers of original music are welcome. No covers. Every 1st and 3rd Monday Night (excluding holidays). 6:30-9:30 PM (Sign up from 6:30pm -8:30pm) Non-smoking and family friendly event. Buffalo West offers delicious entrees, friendly service, and a very nice salad bar, add Live Music and you have a great Date Night! For more information on Buffalo West click HERE> http://www.buffalowestfw.com/


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Smooth-muscle tumors of the gastrointestinal tract are being reported in the current journals with ever-increasing frequency. A review of the world's literature in 1941 contained 972 cases, benign and malignant,7 and many additional reports have been added since. With improvement in diagnostic techniques, it seems reasonable to assume that this rise will continue, and, concomitantly, there will be a larger number of patients with the lesion requiring surgical management. It is becoming necessary, therefore, that surgeons be familiar not only with the pathology and diagnosis of smoothmuscle tumors, but also with proper operative therapy.
In all respects, the management of this lesion is a difficult one. It is unusual for the diagnosis to be made preoperatively, especially with gastric lesions, for there is no characteristic x-ray appearance,4 and the clinical impression is usually that of carcinoma. In the present series of 18 cases, the correct preoperative diagnosis was
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Alley Oop by Jack and Carole Bender
- January 25, 2013
- From Beginning
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Comments (37) (Please sign in to comment)
Nice test, Wizer. :)
Hugpouncies, Oopsters!
Alley Oop: "You called?"
This is gonna get real silly real quick…
he should try "D'Antoni is perfect coach for the lakers", almost no one will say that
Would be kind of funny if that white rabbit ran into a rabbit hole and Wiz and the T-Rex followed him – and they all ended up in "Wonderland!"
Alternate Alley Oop Chapter 16 "Earth 2 Crisis"(56) 1/25/2013
Panel One
"I would suggest we address this device in order to leave." The G.O. Commander tells them.
Panel Two
So several of them as best they can ask in unison to be let out. The floating ball vibrated then its skin shot out like spines about one foot then it went back to what it was previously then it spoke.
[[All of you can leave, stand within the radius of the plate.]] It said in their minds. Plate? They looked and noticed for the first time in the center of the 'village' darker and shiny circular plate to stand on.They all stood within then the next thing that happens is…
Now if the creature stops on command he has what he needs to build up an army.
he should try "D'Antoni is perfect coach for the lakers", almost no one will say that
Not to worry, what could possibly go wrong with this scheme?
This is gonna get real silly real quick…
- – - – - – - – -
"Gonna?" Wadaya mean "gonna?"
This is gonna get real silly real quick…
What do you mean "gonna get"?
Now, what would have been funny is: if the "hypnotic powder" had blow back into Wizer's face when he tried to dose the T-Rex and Wizer had come under the control of the T-Rex!! But that would be too far out for the Benders.
Wootietoot: This is gonna get real silly real quick…
- – – – – – – – -
davidf42:"Gonna?" Wadaya mean "gonna?"
I think it may have gone past silly
Would you think "dumb"?
A great morning to the Benders and all. I guess it wabbit hunting season. WIzer will have to have a megaphone
as a charging T-Rex cannot hear verbal commands very
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Harley Quinn's Greatest Moments from "Batman: The Animated Series"
TV, Comic Books
Comic Books, TV
Film, Comic Books
Comic Books, Film
The Buy Pile
The Buy Pile
The Buy Pile
Robot 6
Robot 6
Comics Should be Good
Spinoff Online
Spinoff Online
Robot 6
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Welcome to Food or Comics?, where every week we talk about what comics we'd buy on Wednesday based on certain spending limits — $15 and $30 — as well as what we'd get if we had extra money or a gift card to spend on a "Splurge" item.
Check out Diamond's release list or ComicList if you'd like to play along in our comments section.
Chris Mautner
If I had $15:
I have a couple of options here. The new issue of The Boys is out ($3.99), as is Vol. 4 of Bakuman ($9.99) and both are currently on my "must-buy" list. But then there's I Will Bite You ($14), a new collection of comics by Joseph Lambert, courtesy of Secret Acres. I've enjoyed the few mini-comics by Lambert that I've read, enough to at least consider putting my other purchases aside in order to get this book instead. There's also what I believe to be the final issue of Alan Moore's Dodgem Logic ($8), which I'd likely ask my retailer to put aside for me for a week when the pickings were slimmer.
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Today's Events
Third Annual Jewish Film Festival at Jewish Community Center
The Midnight Orchestra, the story of the son of a once famous Jewish musician, Marcel Botbol. Directed by Jérôme Cohen Olivar.
Santa Fe Community Farm Stand at Santa Fe Community Farm
Los Radiators • acoustic folk, blues at O'Neill's HeightsMore Recommended Events ››
Kombat Kitty at Stoneface • 7.29.2016
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Push Wins 3 Prizes at Sundance
Last week we mentioned that the novel Push, written by Harlem schoolteacher and poet Sapphire and a popular multi-cultural reading assignment hereabouts, had been made into a film and was debuting at the Sundance Festival. Last night the film won the Festival's jury and audience awards in the U.S. drama category, and a special jury prize for Mo'Nique as the teen heroine's abusive mother. The New York Post's Lou Lumenick described Push: Based on a Novel by Sapphire as "roughly a Harlem version of The Color Purple minus Steven Spielberg's schmaltz," and calls Mariah Carey's performance as a social worker (!) "very well played." Blogging Sundance says the premise "is so unsettling and bleak that no one would blame you if you didn't want to see it," but a "feeling of hopefulness, not the awfulness that precedes it, is what you'll take with you when the film is over." The film has no U.S. release date, but should get one quickly now.
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Installed new stereo in 2008 mitsubishi eclipse gs and now dimmer switch for dash light isnt working but lights come on, except I have no center console light, glove box light, or passenger L.E.D. Tail light but works when I apply the brakes and they are the same L.E.D. Panel
Monday, May 16th, 2011 AT 2:38 AM


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The main objective of Horizon2020 ENERWATER project is to develop, validate and disseminate an innovative standard methodology for continuously assessing, labelling and improving the overall energy performance of Wastewater Treatment Plants (WWTPs).
For that purpose a collaboration framework in the water treatment sector including research groups, SMEs, water management companies, city councils, water authorities and industry was set up.
ENERWATER is devoting important efforts to ensure that the methods are widely adopted. Subsequent objectives are to impulse dialogue towards the creation of a specific European legislation following the example of recently approved EU directives, to establish a way forward to achieve EU energy reductions objectives for 2020, ensuring effluent water quality, environmental protection and compliance with the Water Framework Directive (FWD).
The project will last 36 months starting in 1st March 2015.


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This research was undertaken because of strikingly different experiences with sodium benzoate when taken by mouth in the following ways: (a) pure, as dry crystalline salt, or in aqueous solution; (b) in neutral or alkaline solutions, or in mixtures rich in fat, carbohydrate or protein, e. g., milk; (c) in acid vegetables or fruit, either warm, as in tomato soup, or cold, as in canned plums, peaches, tomatoes, etc.; (d) in beverages containing large percentages of organic acids, e. g., cider, lemonade, grape juice, etc.; (e) in mixtures containing inorganic acids, e. g., artificial gastric juice.
Brunton1 has compiled data that show the comparative arresting influence of various drugs on the action of ferments (Table 1). From these data it is evident that the inhibiting action of a given amount of benzoic acid in combined form, e. g., sodium benzoate, is very much weaker than when the benzoic acid is free.2
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Hotels near Mangala Stadium
Search hotels near Mangala Stadium in Mangalore
Trending now in Mangalore
Hotels within 0.5 miles of Mangala Stadium
- Most bookedHotel Deepa Comforts
4 people looked at this hotel in the last hour
Most recent review
"Our stay was very good. The complimentary breakfast was one of the best I've had. Hotel was clean and the room was spacious and comfortable for the price. People knew good English which was good. Hotel's proximity to city area is a plus. Overall, it's ok for the money you spend."
A Traveller, Nov 2015, US
Hotels within 1 mile of Mangala Stadium
29 people looked at this hotel in the last hour
Most recent review
"At first while booking I was not sure about this place because of other review, but it was the best stay ever. Everything is good about this hotel and its really clean in and outside. I didnt see any other hotel at Mangalore that was this nice. When i come back, I will 100% stay there again."
A Traveller, Jan 2016, GB
Hotels within 2 miles of Mangala Stadium
17 people looked at this hotel in the last hour
Most recent review
"Check out took a little longer than expected. Looks like the reservation and payment details were not easily accessible on their system. As always, have your reservation confirmation details printed out for most hotels in India."
A Traveller, Oct 2015, IN
11 people looked at this hotel in the last hour
Most recent review
"Hotel staff are great. Very friendly and willing to help. Hotel is ok. I stay here all the time while in Mangalore. The price is very fair. Very easy with everything there. Food options are good. Only issue is that they allow customers to smoke inside the rooms, which is fine unless you are like ..."
A Traveller, Mar 2015, US
8 people looked at this hotel in the last hour
6 people looked at this hotel in the last hour
Most recent review
"Hotel is on Mother Theresa Road 2 minutes walk from centre of city. Good hotel, free breakfast n wifi. NOTE, very low balustrade in corridors and windows are wooden frame single glass so careful where you lean and walk. I would definetly stay here again if I were to revisit city."
A Traveller, Oct 2015, GB
4 people looked at this hotel in the last hour
Most recent review
"Nice hotel, a little away from city centre. Very easily accessible from the Highway."
A Traveller, Nov 2015, GB
4 people looked at this hotel in the last hour
- OYO Rooms Jyoti CircleNo Guest Reviews yet
- OYO Rooms Kottara MangaloreNo Guest Reviews yet
Hotels in Mangalore near Mangala Stadium
Are you looking for a cheap Mangala Stadium hotel, a 5 star Mangala Stadium hotel or a family friendly Mangala Stadium hotel? You just landed in the best site to find the best deals and offers on the most amazing accommodations for your stay.
When you search for hotels near Mangala Stadium with Hotels.com, you need to first check our online map and see the distance you will be from Mangala Stadium, India.
Our maps are based on hotel search and display areas and neighborhoods of each hotel so you can see how close you are from Mangala Stadium and refine your search within Mangalore or India based on closest public transportation, restaurants and entertainment so you can easily get around the city. All the hotels details page show an option for free or paid onsite parking.
If you wish to see the hotels with the highest featuring discounts and deals near Mangala Stadium, simply filter by price/ average nightly rate. We recommend you filter by star rating and read our genuine guest reviews so you can get the best quality hotel with the best discount.
One of the new features on Hotels.com guest reviews is that also show reviews from Expedia for Mangala Stadium hotels and the TripAdvisor Mangala Stadium hotels reviews so you can make sure that you checking with a reliable source. See the review scores on our Mangalore hotel information pages.
Make the most out of your family vacation when you book your accommodation with Hotels.com – book your hotel near Mangala Stadium, Mangalore after reviewing the facilities and amenities listed for each hotel.
After booking your hotel near Mangala Stadium, expect to receive your reservation confirmation in the mail in less than 10 minutes. The confirmation email contains more information on all nearby attractions, local directions and weather forecast, so you can better plan the days during your trip.
After getting the best hotel rates you can still save more by winning 1 free night! That's right, book 10 nights in any hotel near Mangala Stadium, Mangalore and after you sign up for the Welcome Rewards program, you are eligible hotel you receive 1 night free*
The best hotel deals are here: We have Mangala Stadium hotel deals, Mangala Stadium last minute deals and offers to get you the cheapest Mangala Stadium hotel with our lowest price guarantee.