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Low temperature incubation improves the performance of anti-phenolic glycolipid-I antibody detecting ELISA in leprosy patients.

Leprosy, caused by Mycobacterium leprae, is a devastating human disease where damage to the peripheral nerves leads to deformities of the hand, foot, face and in some cases eye. A strategy to diagnose leprosy at an early stage and subsequent treatment could help in preventing both serious disability in the patient and transmission, as untreated leprosy is contagious. As of now, accurate and efficient diagnosis of leprosy is considered as a great challenge for leprosy health care. Employing the skin smear test, leprosy patients are classified as smear-positive (for acid-fast bacilli) multibacillary (MB) and smear-negative (for acid-fast bacilli) paucibacillary (PB) types (World Health Organization, 1988). Regarding serology in leprosy, phenolic glycolipid-I (PGL-I) is a highly specific antigen of M. leprae (Hunter & Brennan, 1981) and is known to give rise, predominantly, to IgM antibodies against its terminal trisaccharide. Using PGL-I based antibody detecting tests, well over 90 % of MB patients have high levels of antibodies whereas 15–40 % of the PB patients have detectable antibodies (Buhrer et al., 1998; Chanteau et al., 1989, 1993; Cho et al., 1983, 1991; Fujiwara et al., 1984; Hussain et al., 1990; Roche et al., 1991; Saad et al., 1990). Thus, the major limitation of the serological assays has remained the detection of a limited number of smear-negative PB patients that could be due to an undetectable weak antibody response to M. leprae. Hence, a strategy to improve the detection rate for such patients is clearly needed. In the present study we have investigated the effect of incubating the reactants at low temperature in order to enhance the performance for detecting antibodies to phenolic glycolipid-I antigen.