青藏高原牦牛多杀性巴氏杆菌荧光定量RQ-PCR检测方法的建立  

Development of the real-time fluorescence quantitative RQ-PCR detection assay for Pasteurella multocida of yak in Tibetan plateau

作　　者：赵静[1,2] 张红见[2] 马梅琴[3] 廖明[1] 陈刚[2] 

机构地区：[1]华南农业大学兽医学院人兽共患病防控制剂国家地方联合工程实验室,广东广州510642 [2]青海大学农牧学院,青海西宁810016 [3]青海大学昆仑学院,青海西宁810600

出　　处：《中国兽医杂志》2013年第10期18-20,24,共4页Chinese Journal of Veterinary Medicine

基　　金：国家自然科学基金项目(30960287)

摘　　要：参照GenBank中已发表的多杀性巴氏杆菌种特异性基因KMT-1的保守序列设计1对引物,建立了牦牛多杀性巴氏杆菌实时荧光定量RQ-PCR检测方法。获得的标准曲线相关系数R值为0.9998,标准曲线的线性范围达到1.0×100～1×1010拷贝。对大肠埃希氏菌、肺炎克雷伯氏菌和伤寒沙门菌等非巴氏杆菌进行检测,结果均为阴性。结果表明,本研究建立的荧光定量RQ-PCR方法敏感性高,特异性强,为青藏高原牦牛多杀性巴氏杆菌病的早期诊断及分子流行病学调查提供了一种新的快速检测方法。The primers were designed according to Pasteurella multocida （Pm） species-specific KMT-1 gene conserved sequence from the GenBank and a real time fluorescence RQ-PCR quick detection method of Pm from yak was established. Analysis of standard curves revealed excellent correlation between the number of copies（in the range of 1.0×1 0^0～1×10^10）and PCR threshold cycle（Ct）with a correlation co-efficient of 0. 9998. It was negative to Klebsiella pneumoniae ,Esherichia coli and Salmonella typhi. The results showed high sensitivity and strong specificity, and a new rapid detection methods was established for the early diagnosis and molecular epidemiological investigation of Haemorrhagic cepticaemia of yak in Tibet plateau.

关 键 词：牦牛 多杀性巴氏杆菌 SYBR Green Ⅰ 实时荧光定量RQ-PCR 

分 类 号：S852.615[农业科学—基础兽医学]