动物医学进展2017，Vol.38Issue(3)：59-63,5.
产气荚膜梭菌毒素基因分型PCR检测方法的建立及初步应用
Establishment and Application of PCR Method for Toxin Gene Typing in Clostridium perfringens
摘要
Abstract
This study established a rapid diagnostic PCR method for Clostridium perfringens,provided an effective technical means for the rapid diagnosis and epidemiological investigation of animal Clostridium perfringens infection in order to overcome the long time and high cost disadvantages in traditional identification methods,improve the detection efficiency.Based on the analysis of alpha toxin,beta toxin,epsilon toxin and iota toxin gene sequences for Clostridium perfringens,five pairs of specific primers were designed and synthesized by Premier5.0 software,the PCR method of differential diagnosis for Clostridium perfringens types was established.The optimum annealing temperature was 53 ℃ by repeated experiments.The sensitivity test showed that the PCR can detect the lowest concentrations of DNA of the alpha toxin for 308 pg/L,beta toxin,epsilon toxin for 30.8 pg/L,iota toxin A for 0.122 pg/L,iota toxin B for 0.05 pg/L.The specificity test showed that the method has higher specificity.At the same time,through 16 S rRNA sequence analysis of the positive samples,and compared with GenBank in other Clostridium perfringens 16 S rRNA sequence,the homologies were above 98%.Thus,the established detection method has high sensitivity,strong specificity,and can be used in laboratory diagnosis of Clostridium perfringens infections in animals.关键词
产气荚膜梭菌/毒素基因/聚合酶链反应/鉴别诊断Key words
Clostridium perfringens/toxin gene/PCR/differential diagnosis分类
农业科技引用本文复制引用
赵凤菊,关淼,李井春,杨本勇,赵晓彤..产气荚膜梭菌毒素基因分型PCR检测方法的建立及初步应用[J].动物医学进展,2017,38(3):59-63,5.基金项目
辽宁省农业攻关及产业化项目(2015202013) （2015202013）
