PCR检测兔支气管败血波氏杆菌方法的建立及应用

发布时间：2018-04-29 09:59

本文选题：兔波氏杆菌 + PCR　；参考：《河南农业科学》2017年04期

【摘要】：为建立准确、快速检测兔支气管败血波氏杆菌(Bordetella bronchiseptica)的PCR方法,根据Gen Bank中波氏杆菌ptxA基因设计1对特异性引物,用建立的PCR方法扩增兔波氏杆菌,可扩增出870 bp的目的片段。该方法特异性好,仅对波氏杆菌有特异性的扩增,而对大肠埃希菌、无乳链球菌、金黄色葡萄球菌、多杀性巴氏杆菌等病原菌的扩增结果呈阴性;敏感性较高,最低检测限可达6.8×10~(-7)μg/m L;用建立的PCR方法检测临床疑似感染支气管败血波氏杆菌病兔的脏器及鼻拭子样品,可扩增出目的条带,且与细菌分离的结果一致。可见,建立的检测方法敏感、特异,可用于临床上兔波氏杆菌病的快速诊断及鉴定。
[Abstract]:In order to establish a PCR method for rapid detection of Pasteurella bronchiolus (Bordetella bronchiseptica), 1 pairs of specific primers were designed according to the ptxA gene of Pasteurella in Gen Bank. The amplified fragment of Pasteurella rabbit was amplified by the established PCR method, and the target fragment of 870 BP was amplified. This method has a good specificity and only specific expansion of Pasteurella. The amplification results of Escherichia coli, Streptococcus free Streptococcus, Staphylococcus aureus and Pasteurella multocida were negative, and the sensitivity was higher, the minimum detection limit was 6.8 * 10~ (-7) g/m L, and the PCR method was used to detect the viscera and nose swab samples of clinically suspected bronchobacillosis rabbits. The results showed that the established method is sensitive and specific, and can be used for the rapid diagnosis and identification of Bordetella in rabbits.

【作者单位】：信阳市动物疫病预防控制中心;信阳师范学院生命科学学院;
【基金】：河南省高等学校青年骨干教师资助计划项目(2015GGJS-139)
【分类号】：S852.61

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