Luo Qingping, Huang Hongliang, Zou Wei, Dan Hanbing, Guo Xuebo, Zhang Anding, Yu Zhengjun, Chen Huanchun, Jin Meilin
Unit of Animal Infectious Diseases, National Key Laboratory of Agricultural Microbiology, Huazhong Agricultural University. 1 Shizishan Street, Wuhan, Hubei 430070, PR China.
Vet Microbiol. 2009 May 28;137(1-2):24-30. doi: 10.1016/j.vetmic.2008.12.009. Epub 2008 Dec 13.
A sandwich ELISA test using AIV H5 subtype specific monoclonal antibody (clone 2H4) to an epitope of hemagglutinin protein has been developed. The monoclonal antibody was used to capture the antigen from clinical samples (swabs and tissues). Captured antigens from clinical samples were detected using polyclonal sera, purified AIV H5N1 particles were titrated in the sandwich ELISA and the limit of detection was determined to be approximately 1.0 ng of influenza viral protein in virus preparations. Fifteen AIV strains of H1-H15 subtypes and some other pathogens were tested by this system, and the test is specific to H5 subtype viruses as it failed to detect other AIV subtype viruses and other pathogens. Varieties of clinical samples originating from laboratory experiments (n=382) and from fields (n=288) were employed to test the efficacy of DAS-ELISA test. The test compared very well with the traditional method for detection of influenza virus: virus isolation (VI) in embryonated chicken eggs. In comparison to virus isolation the sensitivity and specificity of sandwich ELISA were found to be 98.6% and 97.6% respectively. In addition, the DAS-ELISA was used to test samples of experimentally infected birds and clinical samples obtained from central China in 2005. The assay proved to be sensitive and specific for the rapid detection of AIV H5 subtype virus form the tissues and swabs in infected animals.
已开发出一种夹心酶联免疫吸附测定（ELISA）试验，该试验使用针对血凝素蛋白表位的禽流感病毒H5亚型特异性单克隆抗体（克隆2H4）。该单克隆抗体用于从临床样本（拭子和组织）中捕获抗原。使用多克隆血清检测临床样本中捕获的抗原，在夹心ELISA中对纯化的禽流感病毒H5N1颗粒进行滴定，确定病毒制剂中流感病毒蛋白的检测限约为1.0纳克。该系统对15种H1 - H15亚型禽流感病毒株和其他一些病原体进行了检测，该试验对H5亚型病毒具有特异性，因为它未能检测到其他禽流感病毒亚型和其他病原体。采用源自实验室实验（n = 382）和现场（n = 288）的各种临床样本，以测试双抗夹心ELISA试验的效果。该试验与检测流感病毒的传统方法：鸡胚病毒分离（VI）相比效果良好。与病毒分离相比，夹心ELISA的灵敏度和特异性分别为98.6%和97.6%。此外，双抗夹心ELISA用于检测2005年实验感染禽类的样本和从中国中部获得的临床样本。该检测方法被证明对从感染动物的组织和拭子中快速检测禽流感病毒H5亚型病毒具有敏感性和特异性。
