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荧光定量PCR技术在霍乱弧菌检测中的应用
引用本文:张世英,洪帮兴,司徒潮满,钟跃星,田华伟,黄慈林,张雪.荧光定量PCR技术在霍乱弧菌检测中的应用[J].中国公共卫生,2003,19(3):345-346.
作者姓名:张世英  洪帮兴  司徒潮满  钟跃星  田华伟  黄慈林  张雪
作者单位:深圳市暨福田区疾病预防控制中心,518033
基金项目:深圳市暨福田区科技三项经费资助
摘    要:目的:建立霍乱弧菌荧光定量聚合酶链反应检测方法,评价其优越性。方法:根据霍乱弧菌NhaA基因保守序列,设计合成霍乱弧菌FQ-PCR诊断试剂盒,比较该方法与常规检测方法在外环境、海产品及临床病例霍乱弧菌检测上的灵敏度和特异性。结果:在临床患标本霍乱弧菌的检测上,FQ-PCR与常规方法灵敏度主、特异性一致,但在含菌量少、菌株易发生变异(外环境疫水、海产品及霍乱越冬)标本的检测上,FQ-PCR显示出其独特的优越性。结论:FQ-PCR方法用于霍乱弧菌的检测,不仅可避免常规PCR引起的假阳性污染,同时可实现对含菌量少、变异菌株的定量定时检测,对临床检验及卫生检测有较大的指导意义。

关 键 词:霍乱弧菌  聚合酶链反应  荧光光度测定法  应用
文章编号:1001-0580(2003)03-0345-02
修稿时间:2002年8月27日

Application of fluorescence quantitative PCR in detection of vibrio cholerae
ZHANG Shi-ying,HONG Bang-xing,SITU Chao-man,et al..Application of fluorescence quantitative PCR in detection of vibrio cholerae[J].Chinese Journal of Public Health,2003,19(3):345-346.
Authors:ZHANG Shi-ying  HONG Bang-xing  SITU Chao-man  
Institution:ZHANG Shi-ying,HONG Bang-xing,SITU Chao-man,et al.Futian Epidemic and Health Prevention Station
Abstract:Objective To construct the detection method of vibrio cholerae with fluorescence quantitative PCR and evaluate its superiority.Methods A diagnostic kit(FQ-PCR) was developed according to the common sequence of NhaA gene in vibrio cholerae.The sensitivity and speciality in detecting of vibrio cholerae in infectious water,sea food and winter-endurance specimen and clinical samples were compared with FQ-PCR kit.Results In the detection of clinical samples,FQ-PCR and regular detection ways have the same sensitivity and speciality.While in the detection of infectious water,sea food and winter-endurance specimen,FQ-PCR has superiority to regular ways.Conclusion The construction of FQ-PCR method eliminates PCR cross-contamination which cause false positive,and real-time detection ensure accurate quantity.So FQ-PCR can be used to the detection of vibrio cholerae in clinical and sanitary laboratory.
Keywords:vibrio cholerae  polymerase chain reaction  fluorophotometry
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