%0 Journal Article
%T Expression of prn gene of Bordetella bronchiseptica and development of a re-combinant protein-based indirect ELISA for antibodies detection<br>猪源支气管败血波氏杆菌百日咳杆菌黏附素基因的原核表达及其抗体检测ELISA方法
%A Zhanqin Zhao
%A Yun Xue
%A Bin Wu
%A Xibiao Tang
%A Huanchun Chen
%A Zengqiang Li
%A Ruiming Hu
%A Jianmin Zhang
%A Longchuan Duan
%A <br>赵战勤
%A 薛云
%A 吴斌
%A 汤细彪
%A 陈焕春
%A 李增强
%A 胡睿铭
%A 张建民
%A 段龙川
%J 微生物学报
%D 2008
%I 
%X OBJECTIVE: We developed an indirect ELISA method for detecting Bordetella bronchiseptica (Bb) pertactin antibodies based on the recombinant pertactin protein expressed in Escherichia coli (DE3) strain. METHODS AND RESULTS: The prn gene encoding Bb pertactin was fused to the downstream of glutathione S-transferase (GST) of pGEX-KG expression vector, resulting in the fusion expression plasmid pGEX-prn. SDS-PAGE showed that the GST-PRN fusion protein was expressed in high level in BL21 carrying pGEX-prn. The strong reactivity of the GST-PRN fusion protein, specifically with antiserum against porcine Bordetellosis caused by Bb HH0809, was identified by Western blot. The recombinant protein fragment of rPRN was purified from the GST-PRN fusion protein digested by protease thrombin with the purity of 93.1%. The rPRN-based indirect ELISA was developed for detecting antibodies against PRN. The ELISA could detect positive samples in experimentally infected pigs fourteen days post inoculation and the degree of sensitivity was over 4 times higher than the latex agglutination test with the coating antigen of killed Bb. Thirty-two point seven percent of positive samples were detected in 1,229 clinical samples while no false positive results were found in detecting 7 antisera against porcine bacterial diseases. Sera samples from two bordetellosis-positive pig fields were tested by the indirect ELISA method and the results indicated that pigs were infected by Bb during the nursery periods. CONCLUSION: The assay showed excellent specificity, sensitivity and reduplication, and can be useful for epidemiological survey and clinical diagnosis of swine bordetellosis.
%K Bordetella bronchiseptica
%K pertactin
%K ELISA<br>支气管败血波氏杆菌
%K 百日咳杆菌黏附素
%K ELISA
%K 猪源
%K 支气管
%K 波氏杆菌
%K 百日咳
%K 黏附素
%K 基因
%K 原核表达
%K 抗体水平
%K 检测阳性率
%K indirect
%K ELISA
%K 方法
%K detection
%K antibodies
%K recombinant
%K development
%K gene
%K 调查
%K 流行病学
%K 氏菌
%K 监测结果
%U http://www.alljournals.cn/get_abstract_url.aspx?pcid=90BA3D13E7F3BC869AC96FB3DA594E3FE34FBF7B8BC0E591&jid=A3C6BA55AB623B90FA9104CFFC826F3C&aid=66793B2308214A05A357B7752D7DCA08&yid=67289AFF6305E306&vid=B6DA1AC076E37400&iid=38B194292C032A66&sid=6C069F4B248105A5&eid=50EA2A80A7D254EF&journal_id=0001-6209&journal_name=微生物学报&referenced_num=0&reference_num=20