title: Development of a specific latex agglutination test based on a recombinant hemagglutinin protein to detect antibodies to H5 avian influenza viruses.
abstract: A latex agglutination test (LAT) was developed for rapid detection of antibodies to H5 avian influenza viruses (AIVs). The hemagglutinin protein of H5 AIV was covalently linked to carboxylated latex by ethyl-dimethyl-amino-propyl carbodiimide to prepare the sensitized latex beads. The LAT was evaluated with the hemagglutination inhibition (HI) assay as the reference test. The H5-LAT showed a sensitivity of 87.0% and specificity of 88.9% in detecting 126 serum samples from experimentally infected chickens and a sensitivity of 82.5% and specificity of 86% in detecting 587 field chicken serum samples from mostly vaccinated chickens. The agreement ratio between H5-LAT and HI was found to be 87.3% and 83.1% for the two groups of samples, respectively. Difficulty with background agglutination in stored chicken sera was overcome by serum pretreatment with either dried chicken liver powder or dilution buffer containing detergent Tween-20. The H5-LAT has advantages over a previously reported whole-virus LAT in terms of biosafety in preparation, chemical stability, and higher specificity. It is a rapid and simple test suitable for field monitoring of antibodies to H5-type AIV.
authors.affiliation: Laboratory of Animal Virology, College of Veterinary Medicine, Unit of Animal Infectious Diseases, Huazhong Agricultural University, Wuhan, People's Republic of China.
authors.forename: Zhengjun
authors.initials: Z
authors.lastname: Yu
authors.forename: Meilin
authors.initials: M
authors.lastname: Jin
authors.forename: Xiaojuan
authors.initials: X
authors.lastname: Xu
authors.forename: Ruihua
authors.initials: R
authors.lastname: Zhang
authors.forename: Hongbo
authors.initials: H
authors.lastname: Zhou
authors.forename: Qiaoyun
authors.initials: Q
authors.lastname: Hu
authors.forename: Haiya
authors.initials: H
authors.lastname: Wu
authors.forename: Bo
authors.initials: B
authors.lastname: Qin
authors.forename: Yad
authors.initials: Y
authors.lastname: Tan
authors.forename: Huanchun
authors.initials: H
authors.lastname: Chen
mesh_terms: D000818:Animals; D000914:Antibodies, Viral / Q000032:analysis*; D000956:Antigens, Viral / Q000032:analysis / Q000276:immunology; D001717:Birds / Q000276:immunology / Q000821:virology; D019267:Hemagglutinin Glycoproteins, Influenza Virus / Q000032:analysis / Q000276:immunology*; D009980:Influenza A virus / Q000276:immunology* / Q000302:isolation & purification; D005585:Influenza in Birds / Q000175:diagnosis* / Q000821:virology; D007841:Latex Fixation Tests / Q000379:methods*; D011994:Recombinant Proteins; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity
chemical_list: D000914:Antibodies, Viral; D000956:Antigens, Viral; D019267:Hemagglutinin Glycoproteins, Influenza Virus; D011994:Recombinant Proteins; C109963:hemagglutinin, avian influenza A virus