羊魏氏梭菌PCR方法的建立及初步应用

›› 2011, Vol. 38 ›› Issue (11): 106-108.

羊魏氏梭菌PCR方法的建立及初步应用

余波¹, 冉懋韬¹, 谭诗文¹, 徐景峨¹, 魏赐开², 艾玉萍¹

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

收稿日期:2011-04-02 修回日期:1900-01-01 出版日期:2011-11-20 发布日期:2011-11-20
通讯作者: 冉懋韬

Establishment and Application of PCR Assay for Clostridium perfringens Isolated from Sheep

YU Bo¹, RAN Mao-tao¹, TAN Shi-wen¹, XU Jing-e¹, WEI Ci-kai², AI Yu-ping¹

1. Guizhou Institute of Animal Husbandry and Veterinary Medicine,Guiyang 550005,China;2. Weng'an County Animal Disease Control Center,Weng'an 550400,China

Received:2011-04-02 Revised:1900-01-01 Online:2011-11-20 Published:2011-11-20

摘要/Abstract

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

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 Escherichia coli,Streptococcus,Staphyloccocus aureus and Pasteurella multocida. 54 clinical samples were detected by PCR.The results revealed that the established PCR assay was sensitive,specific and reproducible and it could be used to detect rapidly Clostridium perfringens isolated from sheep.

Key words: Clostridium perfringens isolated from sheep; alpha-toxin gene; rapid diagnosis

中图分类号:

S858.26

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]. , 2011, 38(11): 106-108.

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]. , 2011, 38(11): 106-108.

[1]	王娟, 李洋洋, 徐光宇, 史蕾, 孙怀昌. 奶牛乳房炎大肠杆菌刃天青微量板快速诊断及药敏试验[J]. , 2016, 43(2): 535-541.
[2]	孙园园;赵鹏. 动物饲料中沙门氏菌环介导等温扩增(LAMP)快速诊断方法的建立[J]. , 2012, 39(1): 32-36.