贝类折光马尔太虫荧光定量PCR检测方法的建立(英文)

目的]建立贝类折光马尔太虫荧光定量PCR检测方法。方法]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。结果]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。结论]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。

Development of a Fluorescence Quantitative PCR Method for Detection of Marteilia refringens in Shellfish

Objective] This paper was to develop a fluorescence quantitative PCR method for detection of M. refringens in shellfish. Method] A pair of primers and a TaqMan probe were designed and synthesized according to the conserved gene sequences of M. refringens in GenBank, so as to develop a fluorescence quantitative PCR method for detection of M. refringens. The developed fluorescence quantitative PCR method was compared with conventional PCR detection. Result] The fluorescence quantitative PCR could detect 40 template copies of plasmid DNA, and its sensitivity was 100 times higher than the conventional PCR. The detection results of Perkinsus sp, Haplosporidium sp, Aeromonas hydrophila, Pseudomonas fluorescens, Vibrio parahaemolyticu, Vibrio alginolyticu, Vibrio rluvialis and Vibrio mimicus were negtive. Conclusion] The fluorescence quantitative PCR method for M. refringens established in this paper is specific, sensitive, rapid and quantitative with good repeatability, which can be used for clinical detection of M. refringens infection.