A monoclonal antibody-based ELISA for differential diagnosis of 2009 pandemic H1N1

作者：Shao, Hongxia^*; Ye, Jianqiang; Vincent, Amy L.; Song, Haichen; Hickman, Danielle; Qin, Aijian; Lamichhane, Chinta; Perez, Daniel R.

来源：Influenza and Other Respiratory Viruses, 2011, 5: 138-142.

• 2009 pandemic H1N1 virus
• ELISA
• mAb

The swine-origin 2009 pandemic H1N1 virus (pdmH1N1) is genetically related to North American swine H1 influenza viruses and unrelated to human seasonal H1 viruses. Currently, specific diagnosis of pdmH1N1 relies on RT-PCR. In order to develop an assay that does not rely in amplification of the viral genome, a conventional sandwich ELISA for detection of the pdmH1N1 was developed. The sandwich ELISA was based on three monoclonal antibodies (3B2, 5H7, and 12F3) against pdmH1N1. 5H7 and 12F3 were selected as capture antibodies and biotin-conjugated 3B2 was subsequently selected as the detection antibody in the ELISA. The results showed the ELISA had high specificity for pdmH1N1 strains and no reaction with other swine H1 viruses, human seasonal H1N1 or H3N2 viruses, or avian influenza viruses. The limit of detection of the ELISA ranged from 3.2 x 10(3) to 1.5 x 10(4) TCID50/ml. When the ELISA was used to detect viruses in nasal wash samples from infected ferrets, it showed 90.1% sensitivity and 100% specificity compared to the "gold standard" - virus isolation. Our studies highlight a convenient assay for specific diagnosis of the 2009 pandemic H1N1-like viruses.

• 出版日期2011-5
• 单位扬州大学