【摘 要】
：
目的:建立树鼩中嗜血杆菌的CODEHOP PCR快速检测方法,为树鼩微生物质量控制提供参考.
方法:应用CODEHOP在线简并引物设计工具,比对NCBI中6株嗜血杆菌的外膜蛋白序列设计
【机 构】
：
中国食品药品检定研究院实验动物资源研究所100050中国医学科学院医学生物学研究所650118
论文部分内容阅读
目的:建立树鼩中嗜血杆菌的CODEHOP PCR快速检测方法,为树鼩微生物质量控制提供参考.
方法:应用CODEHOP在线简并引物设计工具,比对NCBI中6株嗜血杆菌的外膜蛋白序列设计简并引物.通过4株嗜血杆菌标准菌株和24株它属菌株进行特异性和敏感性评价,将建立CODEHOP PCR检测方法,应用于树鼩嗜血杆菌的检测.
结果:简并引物HF-1/HR-8和HF-3/HR3扩增标准菌株副鸡嗜血杆菌(ATCC 29545)、溶血嗜血杆菌(ATCC 33390)、流感嗜血杆菌(ATCC 33391)、副流感嗜血杆菌(ATCC 33392)的目的片段分别为506bp～535bp和344bp～390bp,经测序和NCBI Blast比对,结果准确.两对引物组合,能够有效的区分嗜血杆菌属菌株与它属菌株,检测限最低可达2.1pg/uL.在受试的野生树鼩呼吸道样本中嗜血杆菌阳性率为20/60.
结论:所建立的方法具有较好的特异性和敏感性,操作简便,能够用于动物样品的嗜血杆菌检测.
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