野兔热快速检测方法的建立  

Establishment of Rapid Detection Methods for Tularemia

作　　者：林颖峥 林士佳 张冠楠[2] 李树清[2] 熊炜[2] 郭雨雁 严亚贤[1] Lin Yingzheng;Lin Shijia;Zhang Guannan;Li Shuqing;Xiong Wei;Guo Yuyan;Yan Yaxian(Shanghai Jiaotong University,Shanghai 200240;Shanghai Entry-exit Inspection and Quarantine Bureau,Shanghai 200135)

机构地区：[1]上海交通大学,上海200240 [2]上海出入境检验检疫局,上海200135

出　　处：《中国动物检疫》2017年第10期94-97,共4页China Animal Health Inspection

基　　金：国家重点研发计划项目(2016YFC1202000、2016YFC1202004);上海出入境检验检疫局科研专项(编号HK001-2014)

摘　　要：野兔热是由土拉弗朗西斯菌引起的兔的一种高度接触性、致死性传染病。为加强口岸对进境野生及实验用兔中野兔热的筛查和流行病学调查,基于土拉弗朗西斯菌保守基因序列,建立了PCR和实时PCR检测野兔热的方法。将建立的方法应用于临床样品检测发现经PCR检测,仅有土拉弗朗西斯菌DNA出现条带单一的特异性基因扩增,经实时PCR检测,土拉弗朗西斯菌DNA在10~12个循环处出现显著扩增,对照病毒样品未见扩增;PCR检测土拉弗朗西斯菌DNA模板的下限量为100~10 pg,而实时PCR检测的下限量为100~10 fg。检测结果证实两种检测方法具有良好的特异性、灵敏性和稳定性,适合应用于野兔热的快速检测。Tularemia is a highly contagious and lethal disease caused by Francisella tularensis.In order to investigatethe epidemic character of Tularemia transmission among wild and experimental rabbits,methods of PCR and Real-timePCR to detect Tularemia were established,based on the conserved sequence of Francis tularensis strains.Both methodswere applied to test clinical samples.By PCR detection,only Francis tularensis single band DNA showed bacteria specificgene amplification.By real-time PCR detection,tularemia bacteria DNA in10-12cycles Francis appeared significantamplification,and no amplification was found in the control virus samples.The lower limit of the amount of CRdetected DNA template for tularemia bacteria Francis was100-10pg,and the lower limit of real-time PCR detectionwas100-10fg.The results showed that PCR and Real-time PCR methods were specific,sensitive and reproducible,thus both methods were suitable for Tularemia rapid detection.

关 键 词：野兔热 PCR REAL-TIMEPCR TAQMAN探针 

分 类 号：S852.65[农业科学—基础兽医学]