Development of Epitope-Blocking ELISA for Universal Detection of Antibodies to Human H5N1 Influenza Viruses Export

@article{citeulike:4109346, abstract = {<sec> <title>Background</title> <p>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.</p> </sec><sec> <title>Methodology/Principal Findings</title> <p>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-blocking ELISA for H5N1 were evaluated using chicken antisera to multiple virus clades and other influenza subtypes as well as serum samples from individuals naturally infected with H5N1 or seasonal influenza viruses. The epitope-blocking ELISA results were compared to those of hemagglutinin inhibition (HI) and microneutralization assays. Antibodies to H5N1 were readily detected in immunized animals or convalescent human sera by the epitope-blocking ELISA whereas specimens with antibodies to other influenza subtypes yielded negative results. The assay showed higher sensitivity and specificity as compared to HI and microneutralization.</p> </sec><sec> <title>Conclusions/Significance</title> <p>The epitope-blocking ELISA based on a unique 5F8 mAb provided highly sensitive and 100\% specific detection of antibodies to H5N1 influenza viruses in human sera.</p> </sec>}, author = {Prabakaran, Mookkan and Ho, Hui-Ting and Prabhu, Nayana and Velumani, Sumathy and Szyporta, Milene and He, Fang and Chan, Kwai-Peng and Chen, Li-Mei and Matsuoka, Yumiko and Donis, Ruben O. and Kwang, Jimmy}, citeulike-article-id = {4109346}, citeulike-linkout-0 = {http://dx.doi.org/10.1371/journal.pone.0004566}, day = {24}, doi = {10.1371/journal.pone.0004566}, journal = {PLoS ONE}, keywords = {antibodies, detecting, h5n1, influenza, universal, universality}, month = {February}, number = {2}, pages = {e4566+}, posted-at = {2009-04-27 22:44:09}, priority = {2}, publisher = {Public Library of Science}, title = {Development of Epitope-Blocking ELISA for Universal Detection of Antibodies to Human H5N1 Influenza Viruses}, url = {http://dx.doi.org/10.1371/journal.pone.0004566}, volume = {4}, year = {2009} }

TY - JOUR ID - citeulike:4109346 L3 - citeulike-article-id:4109346 N2 - <sec> <title>Background</title> <p>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.</p> </sec><sec> <title>Methodology/Principal Findings</title> <p>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-blocking ELISA for H5N1 were evaluated using chicken antisera to multiple virus clades and other influenza subtypes as well as serum samples from individuals naturally infected with H5N1 or seasonal influenza viruses. The epitope-blocking ELISA results were compared to those of hemagglutinin inhibition (HI) and microneutralization assays. Antibodies to H5N1 were readily detected in immunized animals or convalescent human sera by the epitope-blocking ELISA whereas specimens with antibodies to other influenza subtypes yielded negative results. The assay showed higher sensitivity and specificity as compared to HI and microneutralization.</p> </sec><sec> <title>Conclusions/Significance</title> <p>The epitope-blocking ELISA based on a unique 5F8 mAb provided highly sensitive and 100% specific detection of antibodies to H5N1 influenza viruses in human sera.</p> </sec> IS - 2 JF - PLoS ONE TI - Development of Epitope-Blocking ELISA for Universal Detection of Antibodies to Human H5N1 Influenza Viruses PB - Public Library of Science VL - 4 SP - e4566 KW - antibodies KW - detecting KW - h5n1 KW - influenza KW - universal KW - universality AU - Prabakaran, Mookkan AU - Ho, Hui-Ting AU - Prabhu, Nayana AU - Velumani, Sumathy AU - Szyporta, Milene AU - He, Fang AU - Chan, Kwai-Peng AU - Chen, Li-Mei AU - Matsuoka, Yumiko AU - Donis, Ruben AU - Kwang, Jimmy PY - 2009/02/24/ UR - http://dx.doi.org/10.1371/journal.pone.0004566 ER -