[1]侯兵兵,陈昌国,李 娜,等.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(01):40-43.[doi:10.3969/j.issn.1671-7414.2018.01.001]
　HOU Bing-bing,CHEN Chang-guo,LI Na,et al.Establishment of a Method for the Identification of Vibrio Parahaemolyticus by Taqman-Probe Fluorescence Quantitative PCR Analysis[J].Journal of Modern Laboratory Medicine,2018,33(01):40-43.[doi:10.3969/j.issn.1671-7414.2018.01.001]
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Taqman-探针荧光定量PCR鉴定副溶血弧菌方法的建立()
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《现代检验医学杂志》[ISSN:/CN:]

卷:
第33卷
期数:
2018年01期
页码:
40-43
栏目:
论著
出版日期:
2018-02-05

文章信息/Info

Title:
Establishment of a Method for the Identification of Vibrio Parahaemolyticus by Taqman-Probe Fluorescence Quantitative PCR Analysis
文章编号:
1671-7414(2018)01-040-04
作者:
侯兵兵; 陈昌国; 李 娜; 赵强元; 陈秋圆; 刘新萍; 董优优
中国人民解放军海军总医院检验科,北京 100048
Author(s):
HOU Bing-bing; CHEN Chang-guo; LI Na;  ZHAO Qiang-yuan; CHEN Qiu-yuan; LIU Xin-ping; DONG You-you
Department of Clinical Laboratory Navy General Hospital PLA,Beijing 100048,China
关键词:
Taqman-探针;  荧光定量PCR;  副溶血弧菌;  toxR基因
分类号:
R503; R378.3
DOI:
10.3969/j.issn.1671-7414.2018.01.001
文献标志码:
A
摘要:
目的 建立一种Taqman荧光定量PCR鉴定副溶血弧菌的方法。方法 以副溶血弧菌标准株(ATCC-VPJS421)和其它常见致病菌标准株为研究对象,通过Gene Bank获取toxR基因的序列,采用生物信息学软件设计特异性PCR引物及Taqman探针,在SLAN 96P荧光定量PCR仪进行扩增检测,评价该检测方法的特异度和灵敏度。结果 ①设计的引物能够扩增出特异性条带; ②扩增体系中0.5 μl探针的扩增效果优于1.0 μl探针; ③Taqman-探针荧光定量PCR检测方法对副溶血弧菌toxR基因的检测灵敏度为10-1 mg/L; ④在检测粪肠球菌、金黄色葡萄球菌、腐生葡萄球菌、霍氏肠杆菌、铜绿假单胞菌、大肠埃希菌、溶藻弧菌、创伤弧菌、梅氏弧菌和弗尼斯弧菌等10种常见致病菌时未出现阳性扩增,特异度为100%。结论 成功建立Taqman探针荧光定量PCR鉴定副溶血弧菌的方法,该方法特异度、灵敏度均较好,适用于副溶血弧菌的快速检测,具有良好的应用价值。
Abstract:
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-softwareto design specific PCR primers and Taqman probe of Vibrio parahaemolyticustoxR gene and detected by fluorescence quantitative PCR instrument.Results ①The designed primers could amplify specific bands.②Theamplification efficiency of the 0.5 μl probe in the amplification system wasbetter than that of the 1.0 μl probe.③The detection sensitivity of toxR geneof Vibrio parahaemolyticus by Taqman fluorescence quantitative PCR was 10-1 mg/L.④The detection method did not show positive amplification in detection of Enterococcus faecalis,Staphylococcus aureus,Saprophytic staphylococcus,Enterobacter hormaechei,Pseudomonas aeruginosa,Escherichia coli,Vibrio alginolyticus,Vibrio vulnficus,Vibrio metschnikovii and Vibrio 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 Vibrio parahaemolyticus and has a good application value.

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备注/Memo

备注/Memo:
作者简介:侯兵兵(1992-),男,大学本科,技师,主要从事病原微生物快速检测方法研究,E-mail:798557114@qq.com。
更新日期/Last Update: 2018-02-07