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目的建立快速检测食品中沙门菌的多重PCR方法。方法应用软件设计4对沙门菌群特异引物,优化多重PCR扩增条件与样品处理方法,分别对61株沙门菌、14株非沙门菌及模拟标本进行检测,观察方法的特异性、敏感性和可行性。结果建立的多重PCR检测所有的沙门菌均能扩出清晰、特异的预期条带,而非沙门菌均未检出;每反应的灵敏度平均为100cfu。对食品中人工污染的沙门菌的检测,最低检测量是10cfu,检测时间为8～9h,与培养法比较,阳性符合率100%;对市售的481份禽制品进行检测,阳性结果12份,而细菌培养法检测阳性结果为10份。结论多重PCR检测沙门菌敏感、特异、省时、省力,适用于沙门菌快速检测。
Objective To establish a multiplex PCR method for rapid detection of Salmonella in food. Methods Four pairs of Salmonella specific primers were designed and used to optimize multiplex PCR amplification conditions and sample processing methods. 61 strains of Salmonella, 14 strains of non-Salmonella and mock samples were tested respectively to observe the specificity, sensitivity and feasibility. Results The multiplex PCR assay established that all Salmonella strains could amplify clear and specific expected bands, but not Salmonella. The sensitivity of each reaction was 100 cfu on average. The detection of artificially contaminated Salmonella in food was 10cfu, the detection time was 8 ~ 9h, compared with the culture method, the positive coincidence rate was 100%; 481 commercially available poultry products were tested, and the positive result was 12 , While the bacterial culture test positive for 10 results. Conclusion Multiplex PCR detection of Salmonella sensitive, specific, time-saving and labor-saving, suitable for rapid detection of Salmonella.