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Development and diagnostic application/evaluation of pandemic (H1N1) 2009 influenza virus‐specific monoclonal antibodies

We prepared mAb specific to the H1N1 2009 virus (H1N1 2009) to facilitate development of an RDT with enhanced sensitivity and specificity. Among these antibodies, we identified two clones—hybridomas 1H7E1 and 3A3H7—that specifically bound to H1N1 2009 (non‐seasonal) and were very suitable for application to a diagnostic kit. The affinity constants (Ka) of 1H7E1 and 3A3H7 were 1.10 × 1010 and 2.35 × 1010, respectively. To identify the antibodies, we performed ELISA and immunoblot analyses and found that 1H7E1 recognized a conformational epitope of HA while 3A3H7 recognized a linear epitope. In clinical evaluations using specimens from 215 patients, a lateral flow rapid testing kit comprising these mAb showed a sensitivity of 81.5% (75/92) and a specificity of 96.7% (119/123). Results using the RDT kit were well correlated with conventional RT‐PCR methods as commonly and commercially used. Based on our findings, we believe that use of these mAb with a rapid evaluation kit could serve as a good diagnostic tool for H1N1 2009.