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羊魏氏梭菌PCR方法的建立及初步应用
引用本文:余波,冉懋韬,谭诗文,徐景峨,魏赐开,艾玉萍.羊魏氏梭菌PCR方法的建立及初步应用[J].中国畜牧兽医,2011,38(11):106-108.
作者姓名:余波  冉懋韬  谭诗文  徐景峨  魏赐开  艾玉萍
作者单位:1. 贵州省畜牧兽医研究所,贵州贵阳 550005;2. 贵州省瓮安县动物预控中心,贵州瓮安 550400
基金项目:贵州省农业科技攻关项目[黔科合NY字(2009)]3089号]; 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号]; 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]; 贵州省农科院专项[黔农科院院专项(2010)047号]
摘    要:根据羊魏氏梭菌A、B、C、D、E型共有的α毒素基因,设计了1对引物,通过对PCR反应条件进行优化,建立了羊魏氏梭菌PCR检测方法。该方法扩增条带大小为255 bp,最低核酸检测量A型为0.39 ng/L、B型为0.62 ng/L、C型为0.52 ng/L、D型为0.87 ng/L、E型为0.92 ng/L,而对羊大肠杆菌、羊链球菌、金黄色葡萄球菌、羊多杀性巴氏杆菌的扩增结果均为阴性。应用该PCR方法对54份临床样本进行检测,PCR检测结果高于细菌学和生化检测结果。结果表明,该PCR方法具有很好的特异性和敏感性,可用于临床羊魏氏梭菌病的早期快速诊断。

关 键 词:羊魏氏梭菌  α毒素基因  快速诊断  
收稿时间:2011-04-02

Establishment and Application of PCR Assay for Clostridium perfringens Isolated from Sheep
YU Bo,RAN Mao-tao,TAN Shi-wen,XU Jing-e,WEI Ci-kai,AI Yu-ping.Establishment and Application of PCR Assay for Clostridium perfringens Isolated from Sheep[J].China Animal Husbandry & Veterinary Medicine,2011,38(11):106-108.
Authors:YU Bo  RAN Mao-tao  TAN Shi-wen  XU Jing-e  WEI Ci-kai  AI Yu-ping
Institution:1. Guizhou Institute of Animal Husbandry and Veterinary Medicine,Guiyang 550005,China;2. Weng'an County Animal Disease Control Center,Weng'an 550400,China
Abstract:According to the gene sequences in GenBank of alpha-toxin,one pair of specific primer was designed for amplifying the specific fragments of alpha-toxin gene.After optimization of annealing temperature and primers concentrations,PCR assay was established for simultaneous detection of the Clostridium perfringens.The specific bands of 255 bp was amplified.The sensitivity and specificity tests showed that the PCR were sensitive in 0.39,0.62,0.52,0.87,0.92 ng/L A,B,C,D,EC.perfringens.No band was amplified from E...
Keywords:Clostridium perfringens isolated from sheep  alpha-toxin gene  rapid diagnosis  
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