| 检测禽流感病毒抗体的重组核蛋白间接ELISA方法的建立 |
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| 引用本文： | 吴仁蔚,胡思顺,肖运才,李自力,石德时,毕丁仁.检测禽流感病毒抗体的重组核蛋白间接ELISA方法的建立[J].畜牧兽医学报,2006,37(10):1067-1072. |
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| 作者姓名： | 吴仁蔚 胡思顺 肖运才 李自力 石德时 毕丁仁 |
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| 作者单位： | 华中农业大学动物医学院微生物与免疫学研究室,武汉,430070 |
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| 基金项目： | 湖北省科技厅重点项目（20002P0805） |
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| 摘 要： | 以大肠杆菌系统表达的H9N2亚型禽流感病毒（AIV）核蛋白（NP）为抗原，建立了禽流感间接酶联免疫吸附试验抗体检测技术（NP—ELISA）。对263份待检血清（包括临床收集的243份血清和20份H9N2亚型AIV免疫鸡阳性血清）进行检测，NP—ELISA与琼脂免疫扩散试验（AGP）的总符合率为83．3％，与血凝抑制试验（HI）的总符合率为92％。特异性试验表明，NP—ELISA方法可以检测H5、H7和H9亚型AIV特异性抗体，检测为阳性的血清样品能够被阳性鸡胚尿囊液阻断。敏感性试验证实，NP—ELISA最早可以检测鸡感染后7d的血清样品，并于感染后10d确定100％血清阳性，而AGP检测直到首免后21～28d才出现部分血清阳性，HI检测直到10～14d才出现部分血清阳性，并且NP-ELISA要比HI敏感4～40倍。试验证明，NP—ELISA是检测AIV血清型特异性抗体的一种特异、敏感、快速、经济的血清学检测技术。
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| 关 键 词： | 禽流感病毒 重组核蛋白 NP-ELISA 抗体检测 |
| 文章编号： | 0366-6964（2006）10-1067-06 |
| 收稿时间： | 2005-12-05 |
| 修稿时间： | 2005-12-05 |
Development of Indirect Enzyme-linked Immunosorbent Assay with Nucleoprotein as Antigen for Detecting Antibodies against Avian Influenza Virus |
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| WU Ren-wei,HU Si-shun,XIAO Yun-cai,LI Zi-li,SHI De-shi,BI Ding-ren.Development of Indirect Enzyme-linked Immunosorbent Assay with Nucleoprotein as Antigen for Detecting Antibodies against Avian Influenza Virus[J].Acta Veterinaria et Zootechnica Sinica,2006,37(10):1067-1072. |
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| Authors: | WU Ren-wei HU Si-shun XIAO Yun-cai LI Zi-li SHI De-shi BI Ding-ren |
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| Institution: | Laboratory of Animal Microbiology and Immunology, Huazhong Agricultural University, Wuhan 430070, China |
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| Abstract: | Indirect enzyme-linked immunosorbent assay(ELISA) for detecting antibodies against avian influenza virus(AIV) was developed by using expressed full length nucleoprotein(NP) of H9N2 AIV in E.coli.263 chicken serum samples(including 243 clinical serum samples and 20 positive serum samples from H9N2 AIV vaccinated chicks) were detected by NP-ELISA,agar gel precipitin test(AGP),and hemagglutination inhibition(HI).The results showed that the coincidence ratio between NP-ELISA and AGP or HI was 83.3% and 92% respectively.The specific assay suggested that NP-ELISA was able to detect H5,H7 and H9 subtype antibodies to AIV,and the serum samples which were confirmed positive by NP-ELISA could be blocked by positive chicken-embryo allantoic fluid.The sensitive analysis demonstrated that NP-ELISA can detect specific antibody in the 7th day after AIV infection in chicks,and all sera were positive in the 10th day.However,serum samples were still negative at the 21st day post inoculation(PI) by AGP test,and HI tests began to detect low levels of antibodies at the 10th day PI.The sensitivity of NP-ELISA was 4-40 times higher than that of HI.The present study confirmed that the NP-ELISA was a rapid,sensitive,economic and specific method for type-serologically detection of AIV infection in chickens. |
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| Keywords: | avian influenza virus recombinant nucleoprotein NP-EI ISA detecting antibody |
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