现代检验医学杂志2018，Vol.33Issue(1)：40-43,4.DOI:10.3969/j.issn.1671-7414.2018.01.010
Taqman-探针荧光定量PCR鉴定副溶血弧菌方法的建立
Establishment of a Method for the Identification of Vibrio Parahaemolyticus by Taqman-Probe Fluorescence Quantitative PCR Analysis
侯兵兵 1陈昌国 1李娜 1赵强元 1陈秋圆 1刘新萍 1董优优1
作者信息
- 1. 中国人民解放军海军总医院检验科,北京 100048
- 折叠
摘要
Abstract
Objective To establish a method for the identification of Vibrio parahaemolyticus based on Taqman-fluorescence probe quantitative PCR method targeting toxR gene.Methods Taking the standard strain of Vibrio parahaemolyticus (VPJS421) and other ommon pathogenic bacteria'standard strain as the research object,using the bio-software to design specific PCR primers and Taqman probe of Vibrio parahaemolyticus toxR gene and detected by fluorescence quantitative PCR instrument.Results ①The designed primers could amplify specific bands.②The amplification efficiency of the 0.5 μl probe in the amplification system was better than that of the 1.0μl probe.③The detection sensitivity of toxR gene of Vibrio parahaemolyticus by Taqman fluorescence quantitative PCR was 10-1 mg/L.④The detection method did not show positive amplification in detection of Enterococcus f aecalis,Staphylococcus aureus,Saprophytic staphylococcus,Enterobacter hormaechei,Pseudomonas aeruginosa,Escheri ch ia coli,Vibrio al ginol yticus,Vibrio vulnficus,Vibrio metschnikovii and Wbrio furnissii 10 other common pathogenic bacteria.The specificity was 100％.Conlusion The fluorescence quantitative PCR method for the identification of Vibrio parahaemolyticus was successfully established.The method was sensitivty and specificity,and it is suitable for rapid detection of Wbrio parahaemolyticus and has a good application value.关键词
Taqman-探针/荧光定量PCR/副溶血弧菌/toxR基因Key words
taqman fluorescence probe/fluorescence quantitative PCR/vibrio parahaemolyticus/toxR gene分类
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侯兵兵,陈昌国,李娜,赵强元,陈秋圆,刘新萍,董优优..Taqman-探针荧光定量PCR鉴定副溶血弧菌方法的建立[J].现代检验医学杂志,2018,33(1):40-43,4.
