Establishment of a Method for the Identification of Vibrio Parahaemolyticus by Taqman-Probe Fluorescence Quantitative PCR Analysis
10.3969/j.issn.1671-7414.2018.01.010
- VernacularTitle:Taqman-探针荧光定量PCR鉴定副溶血弧菌方法的建立
- Author:
Bing-Bing HOU
1
;
Chang-Guo CHEN
;
Na LI
;
Qiang-Yuan ZHAO
;
Qiu-Yuan CHEN
;
Xin-Ping LIU
;
You-You DONG
Author Information
1. 中国人民解放军海军总医院检验科
- Keywords:
taqman fluorescence probe;
fluorescence quantitative PCR;
vibrio parahaemolyticus;
toxR gene
- From:
Journal of Modern Laboratory Medicine
2018;33(1):40-43
- CountryChina
- Language:Chinese
-
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.
