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目的建立基于toxR基因的PCR检测副溶血弧菌的方法,并通过实验条件摸索,优化反应体系条件,验证PCR方法检测副溶血弧菌的优势。方法应用基于toxR基因的环引物PCR法和已报道的PCR法对副溶血性弧菌和食品样品进行检测,并优化反应条件。对两种检测方法的灵敏度、特异度和检测时间进行比较。结果环引物PCR法可以使副溶血性弧菌扩增出大小180 bp的toxR基因片段,两种方法检测副溶血性弧菌的特异度均为100%,检出限分别为4×103CFU/mL和5×104CFU/mL,检测所需时间分别为2 h 50 min和3 h10 min。结论基于toxR基因的环引物PCR法检测副溶血性弧菌的特异性更强,检出限更低,检测所需时间更短,能够较好地满足卫生防疫机构快速检测副溶血性弧菌的需要。
Objective To establish a method for PCR detection of Vibrio parahaemolyticus based on toxR gene and explore the conditions of the reaction system through experimental conditions to verify the advantages of PCR in the detection of Vibrio parahaemolyticus. Methods Vibrio parahaemolyticus (Vibrio parahaemolyticus) and food samples were detected by PCR using the toxR gene and PCR. The reaction conditions were optimized. The sensitivity, specificity and detection time of the two methods were compared. The results of ring-primer PCR method to make Vibrio parahaemolyticus amplified toxR gene fragment of 180 bp in size, Vibrio parahaemolyticus detection by both methods were 100% specificity, detection limits were 4 × 103CFU / mL And 5 × 104CFU / mL, the time required for the test was 2 h 50 min and 3 h 10 min, respectively. Conclusion The PCR method based on toxR gene is more specific and sensitive to Vibrio parahaemolyticus with lower detection limit and shorter detection time. It can meet the requirements of rapid detection of Vibrio parahaemolyticus by hygienic and epidemic prevention agencies need.