应用荧光PCR方法检测食品中的单核细胞增生李斯特氏菌

Detection of Listeria Monocytogenes in Food by Fluorescence PCR

[目的]发展一种快速、敏感、特异的荧光PCR方法检测食品中单核细胞增生李斯特氏菌。[方法]针对单增李斯特氏菌溶血素基因(hlyA),设计特异的引物和TaqMan探针,优化体系,建立荧光PCR检测方法,对该方法的特异性和敏感性进行评价,并且在对188个进出口食品样本的检测中与传统方法进行对比。[结果]经对20株不同菌属的标准菌株和30株野生李斯特氏菌菌株检测,结果显示该方法具有良好的特异性。灵敏度检测结果表明,经过24h增菌,该方法的检测低限为1cfu/g,整个流程只需27h。在实际样本的检测中,检测结果与传统方法结果一致。[结论]该方法能快速、简便和准确地检出单核细胞增生李斯氏菌,具有良好的应用前景。

This is to develop a rapid,sensitive and specific fluorescence PCR method detecting listeria monocytogenes in food.One set of primers and fluorescent probe were designed according to the listeriolysin O gene(hly) of listeria monocytogenes.We optimized the condition of PCR reaction,established an adapted fluorescence PCR method for food and studied on the sensitivity and specificity of the FQ-PCR method.Using this method,188 import and export samples were detected,and the results were compared with those by traditional method.The assays were 100% specific,as determined with 30 wild listeria strains and 20 standard strains.The threshold is as low as 1cfu/g with simulative samples after these samples were incubated for 24h.The results were the same as traditional method as well as the method is simple,rapid and accurate with good prosperity