[用于检测抗甲型流感病毒（H1N1）抗体的间接酶联免疫吸附测定法]

[Indirect ELISA for detection of antibodies against swine influenza virus (H1N1)].

作者信息

Gao Lei, Liu Sidang, Xiao Yihong, Liu Weimin, Liu Wenjun, Sun Lei

机构信息

College of Veterinary Medicine, Shandong Agricultural University, Tai'an 271018, China.

出版信息

Sheng Wu Gong Cheng Xue Bao. 2011 May;27(5):805-11.

PMID:21845848

Abstract

In order to detect antibody against swine influenza virus (H1N1), HA1 region of hemagglutinin gene in epidemic swine influenza virus (H1N1) strain was amplified and subcloned into prokaryotic expression vector pET30a. Then recombinant HA1 protein was expressed by Escherichia coli BL21. The purified recombinant HA1 protein was obtained after the treatment of denaturing, refolding and affinity chromatography with immobilized nickel chelating NTA (Ni-NTA). An indirect enzyme-linked immunosorbent assay (ELISA) method was established using the purified protein as antigen. Then 785 swine serum samples collected during 2008-2009 were detected by this method, and the positive ratio was 15.54%. There were diversities among provinces (8%-47%). The diagnostic specificity and diagnostic sensitivity of this method arrived at 91% and 95% respectively, using the results of IDEXX ELISA kit as reference.

摘要

为检测猪流感病毒（H1N1）抗体，扩增流行株猪流感病毒（H1N1）血凝素基因的HA1区，并亚克隆至原核表达载体pET30a。然后用大肠杆菌BL21表达重组HA1蛋白。经变性、复性及固定化镍螯合NTA（Ni-NTA）亲和层析处理后，获得纯化的重组HA1蛋白。以纯化蛋白为抗原建立间接酶联免疫吸附测定（ELISA）方法。然后用该方法检测2008 - 2009年采集的785份猪血清样本，阳性率为15.54%。各省之间存在差异（8% - 47%）。以IDEXX ELISA试剂盒检测结果为参照，该方法的诊断特异性和诊断敏感性分别达到91%和95%。