摘要
根据基因库中折光马尔太虫的基因保守序列,设计合成了一对引物和一条TaqMan探针,建立检测折光马尔太虫的荧光定量PCR方法,将建立的荧光定量PCR检测方法与常规PCR检测对比,所建立的荧光定量PCR方法灵敏度可达40个拷贝/μl,比常规PCR灵敏度高100倍,对派琴虫、单孢子虫、嗜水气单胞菌、荧光假单胞菌、副溶血弧菌、溶藻弧菌、河弧菌和拟态弧菌等病原体的检测结果为阴性。结果显示研究建立的折光马尔太虫荧光定量PCR方法具有特异、敏感、快速、定量、重复性好等优点,可用于临床上折光马尔太虫感染的检测。
A pair of primers and a TaqMan probe were designed and synthesized according to the conserved gene sequences of Marteilia refringens,and then reaction parameters were optimized to develop a real-time TaqMan-quantitative PCR assay.After having compared the developed real-time PCR assay with that of routine PCR,the results showed that this real-time PCR assay could detect 40 template copies of plasmid DNA,and its sensitivity was 100 times higher than that of the routine PCR.This real-time PCR results were negative after detecting the samples such as Perkinsus sp.,Haplosporidium sp.,Aeromonas hydrophila,Pseudomonas fluorescens,Vibrio parahaemolyticu,Vibrio alginolyticu,Vibrio fluvialis and Vibrio mimicus.As a result of the sensitivity and specificity of the assay with a relatively rapid and simple procedure,the real-time PCR will be useful as a routine assay for the clinical diagnosis of Marteilia refringens infection.
出处
《西南农业学报》
CSCD
北大核心
2012年第1期306-309,共4页
Southwest China Journal of Agricultural Sciences
基金
国家百千万人才工程人选专项资金项目(945200603)
广西特聘专家专项经费(2011B020)
广西科技攻关项目(桂科攻0630001-3M)
