Development and evaluation of an indirect enzyme-linked immunosorbent assay for the detection of antibodies against Campylobacter fetus in cattle

文献类型： 外文期刊

第一作者： Zhao, Hailing

作者： Zhao, Hailing;Liu, Huifang;Du, Yanfen;Liu, Siguo;Ni, Hongbo;Wang, Yong;Wang, Chunlai;Si, Wei;Yang, Jinguo;Ling, Jingkai

作者机构：

关键词： Campylobacter fetus;Enzyme-linked immunosorbent assay;Prokaryotic expression;Purification;SapA gene

期刊名称：RESEARCH IN VETERINARY SCIENCE （ 影响因子：2.534； 五年影响因子：2.382 ）

ISSN： 0034-5288

年卷期： 2010 年 88 卷 3 期

页码：

收录情况： SCI

摘要： Campylobacteriosis is a zoonosis that occurs worldwide. Infection with Campylobacter fetus (C. fetus) causes infertility and abortion in sheep and cattle. The current study focuses on the SapA gene of C. fetus that encodes surface array proteins and plays an important role in the virulence of C. fetus. The SapA-N (1398 bp) and SapA-C (1422 bp) fragments were amplified from the C. fetus SapA gene using polymerase chain reaction (PCR), and the corresponding recombinant proteins rSapA-N and rSapA-C were expressed in Escherichia. coli BL21 cells. Results of Western blotting and enzyme-linked immunosorbent assay (ELISA) showed that the immunological activity of rSapA-N was higher than that of rSapA-C (P < 0.05). Therefore, rSapA-N was selected to establish an indirect ELISA for detecting antibodies against C. fetus. The diagnostic criteria were as follows: S/P >= 0.45: positive; S/P < 0.4: negative; 0.45 > S/P >= 0.4: suspected. The specificity and sensitivity of our method were 94.3% and 88.6%, respectively. Moreover, no cross-reactions were observed between rSapA-N and serum samples that were positive for other bovine bacterial pathogens diseases such as Mycobacterium avium subspecies paratuberculosis. One hundred and two serum samples from cows that had experienced abortion were tested. Four and 2 C fetus-positive serum samples were found among the 70 bovine brucellosis-positive samples and the 32 infectious bovine rhinotracheitis (IBR)-positive samples, respectively. The findings suggest that the rSapA-N-based ELISA method has immense potential in future applications. (C) 2009 Elsevier Ltd. All rights reserved.

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