牛结核病PCR检测试剂盒的研制

中国畜牧兽医

牛结核病PCR检测试剂盒的研制

吴位珩¹, 徐景峨¹, 孙启跃¹, 杨莉¹, 文才智², 杨茂生¹

1. 贵州省畜牧兽医研究所,贵州贵阳 550005;
2. 贵州省农业科学院科研处,贵州贵阳 550006

收稿日期:2012-01-06 出版日期:2012-08-20 发布日期:2012-08-24
通讯作者: 杨茂生
作者简介:吴位珩(1966－),女,贵州人,副研究员,主要从事畜禽疾病研究工作。
基金资助:
贵州省农业攻关项目"牛结核病PCR诊断试剂盒的研制与应用"(黔科合NY(2009)3086);贵州省畜禽健康养殖技术创新能力建设项目(黔科合院所创能(2010)4004);中央补助地方科技基础条件专项基金项目(黔科条中补地(2010)4001)。

Establishment and Development of PCR Kit for Detecting Mycobacterium tuberculosis

WU Wei-heng¹, XU Jing-e¹, SUN Qi-yue¹, YANG Li¹, WEN Cai-zi², YANG Mao-sheng¹

1. Guizhou Animal Husbandry and Veterinary Medicine Institute,Guiyang 550005,China;
2. Guizhou Academy of Agricultural Sciences,Guiyang 550006,China

Received:2012-01-06 Online:2012-08-20 Published:2012-08-24

摘要/Abstract

摘要： 根据GenBank中的牛结核分枝杆菌IS6110的基因片段,设计了1对引物,通过对PCR反应条件进行优化,研制了用于检测牛结核病的PCR试剂盒,该试剂盒扩增的阳性条带为317 bp;敏感性结果显示,该PCR检测试剂盒的最低核酸检测量为1.025 pg/μL;特异性试验表明,仅结核分枝杆菌扩增结果为阳性,副结核分枝杆菌、胸膜肺炎放线杆菌、大肠杆菌、巴氏杆菌、沙门氏菌、金葡萄球菌、链球菌的扩增结果均为阴性。-20 ℃至少可保存12个月,且重复性良好。应用该PCR试剂盒对24份临床样本进行了检测,其PCR检测结果与结核菌素试验检测结果相一致。结果表明,牛结核病PCR检测试剂盒能够对牛结核临床样本进行快捷、灵敏、准确的检测。

关键词: 牛结核分枝杆菌; PCR试剂盒; IS6110基因

Abstract: A PCR kit for detection bovine tuberculosis IS6110 gene was developed based on PCR principle with designing a pair of primer from GenBank. A specific band (317 bp) was detected with the PCR kit. By sensitivity analysis,a minimum of 1.025 pg pathogen DNA was detected, and pathogens of control groups,such as Pseudo tuberculosis,Actnobacillus,Escherichia coli,Pastueurella,Salmonella,Staphylococcus aureus,Streptococci showed nagative.The kit could be stored at －20 ℃ for a year and having a repeating performance. With the PCR kit, 24 cases was diagnosed and the result was similar to tuberculin test. The PCR kit can be a good method for clinical diagnosis of bovine tuberculosis with rapid, sensitivity and specificity.

Key words: bovine tuberculosis; PCR kit; IS6110 gene

中图分类号:

S855.2

吴位珩, 徐景峨, 孙启跃, 杨莉, 文才智, 杨茂生. 牛结核病PCR检测试剂盒的研制[J]. 中国畜牧兽医.

WU Wei-heng, XU Jing-e, SUN Qi-yue, YANG Li, WEN Cai-zi, YANG Mao-sheng. Establishment and Development of PCR Kit for Detecting Mycobacterium tuberculosis[J]. .

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