development of epitope-blocking elisa for universal detection of antibodies to human h5n1 influenza viruses开发通用epitope-blocking elisa检测h5n1流感病毒的抗体.pdfVIP

Development of Epitope-Blocking ELISA for Universal Detection of Antibodies to Human H5N1 Influenza Viruses 1 1 1 1 1 1 Mookkan Prabakaran , Hui-Ting Ho , Nayana Prabhu , Sumathy Velumani , Milene Szyporta , Fang He , 2 3 3 3 1,4 Kwai-Peng Chan , Li-Mei Chen , Yumiko Matsuoka , Ruben O. Donis , Jimmy Kwang * 1 Animal Health Biotechnology, Temasek Life Sciences Laboratory, National University of Singapore, Singapore, Singapore, 2 Department of Pathology, Singapore General Hospital, Singapore, Singapore, 3 Influenza Division, Centers for Disease Control and Prevention, Atlanta, Georgia, United States of America, 4 Department of Microbiology, Faculty of Medicine, National University of Singapore, Singapore, Singapore Abstract Background: Human infections with highly pathogenic H5N1 avian influenza viruses have generally been confirmed by molecular amplification or culture-based methods. Serologic surveillance has potential advantages which have not been realized because rapid and specific serologic tests to detect H5N1 infection are not widely available. Methodology/Principal Findings: Here we describe an epitope-blocking ELISA to detect specific antibodies to H5N1 viruses in human or animal sera. The assay relies on a novel monoclonal antibody (5F8) that binds to an epitope comprising amino acid residues 274–281 (CNTKCQTP) in the HA1 region of H5 hemagglutinin. Database search analysis of publicly available sequences revealed that this epitope is conserved in 100% of the 163 H5N1 viruses isolated from humans. The sensitivity and specificity of the epitope-blockin