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目的:用TapMan探针的Real-time PCR技术检测鉴定食品中的副溶血弧菌污染定性,并定量检测试剂盒。方法:以副溶血弧菌gyrB基因序列为依据,设计并合成TapMan探针和特异性引物,将扩增的特异片断制成阳性模板,并绘制标准曲线。结果:通过对8种细菌(10株菌株)的DNA进行扩增,发现3株副溶血弧菌均可产生扩增曲线,而其它的7株非溶血弧菌均不产生扩增曲线。结论:TapMan探针的Real-time PCR技术具有很高的特异性及敏感性,能有效避免污染,值得临床广泛应用及推广。
OBJECTIVE: To detect the qualitative and quantitative detection of Vibrio parahaemolyticus in food by Real-time PCR with TapMan probe. Methods: According to gyrB gene sequence of Vibrio parahaemolyticus, TapMan probe and specific primers were designed and synthesized. The amplified specific fragment was made into a positive template and a standard curve was drawn. Results: By amplifying the DNA of 8 kinds of bacteria (10 strains), it was found that all 3 strains of V. parahaemolyticus could produce amplification curve, while none of the other 7 strains of non-lyso-bacterium did not produce amplification curve. Conclusion: The Real-time PCR technique of TapMan probe has high specificity and sensitivity, which can effectively avoid contamination and deserves wide application and promotion in clinic.