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目的建立双重荧光PCR用于检测食物中毒中副溶血性弧菌和沙门菌。方法设计特异性引物和探针用于扩增副溶血性弧菌tlh基因和沙门菌invA基因(invA),采用倍比稀释法检测方法的DNA灵敏度和菌液灵敏度,采用9株其他肠道致病菌验证方法的特异性。结果两种病原菌DNA检测灵敏度分别为65 fg/PCR和56 fg/PCR体系,菌液检测灵敏度分别为11 cfu/ml和9 cfu/ml,特异度100%。用建立的方法对4起食物中毒进行检测,共检出副溶血性弧菌15株和沙门菌8株。结论该方法高效并具有很高的灵敏性和特异性,在副溶血性弧菌和沙门菌引起的食物中毒的快速检测中具有广阔的应用前景。
Objective To establish a dual-fluorescence PCR for the detection of Vibrio parahaemolyticus and Salmonella in food poisoning. Methods Specific primers and probes were designed to amplify the tlh gene of Vibrio parahaemolyticus and the invA gene of Salmonella (invA). The DNA sensitivity and bacterial sensitivity of the method were evaluated by fold dilution method. Specificity of germ verification methods. Results The detection sensitivity of the two kinds of pathogenic bacteria DNA was 65 fg / PCR and 56 fg / PCR respectively. The detection sensitivity was 11 cfu / ml and 9 cfu / ml respectively and the specificity was 100%. Four food poisoning tests were carried out with the established method, and 15 strains of Vibrio parahaemolyticus and 8 strains of Salmonella were detected. Conclusion The method is highly efficient and has high sensitivity and specificity. It has broad application prospects in the rapid detection of food poisoning caused by Vibrio parahaemolyticus and Salmonella.