荧光定量PCR技术在霍乱弧菌检测中的应用

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。

Application of fluorescence quantitative PCR in detection of vibrio cholerae

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.