荧光PCR法与细菌培养法在食源性致病检测中的应用

Application of FQ-PCR and bacterial culture in food-borne pathogens detection

目的:通过荧光PCR法与细菌培养法在食源性致病菌检测中的应用,比较两种方法的敏感性、特异性、时限性、重复性。方法:2009年1月-2010年12月采集的样品进行常见食源性致病的细菌培养,随机抽取一部分样品进行荧光PCR检测。结果:细菌培养法在257份样品中检出单核细胞增生李斯特菌5株、金黄色葡萄球菌5株,检出率3.9%。所检出致病菌经荧光PCR法符合结果,符合率100%。随机抽取各类产品的20%共计49份样品进行沙门菌、大肠埃希菌O157:H7、单核细胞增生李斯特菌、副溶血弧菌、金黄色葡萄球菌等核酸试剂盒检测。结果检出沙门菌1份、金黄色葡萄球菌2份、单核细胞增生李斯特菌1份、副溶血弧菌1份,其余致病菌核酸检测均未检出,检出率10%。结论:与传统的细菌培养法相比,荧光PCR法可提高食源性致病菌的检出率、缩短检出时间、适用于食源性致病菌的检测。

Objective：Sensitivity,specificity,timeliness and repeatability of FQ-PCR and bacterial culture were compared in the detection of food-borne pathogens.Methods： Common food-borne pathogenic bacterial culture was done by using samples collected from Jan 2009 to Dec 2010,and then some samples were selected randomly to make FQ-PCR detection.Results： 5 Listeria monocytogenes bacteria and 5 Staphylococcus aureus are detected by using bacterial culture among 257 samples,with a detection rate of 3.9%.The pathogens are exactly the same as the results detected by FQ-PCR,and the consistent rate is 100%.Nucleic acid kit detection like Salmonella,E.coli O157： H7,Listeria monocytogenes bacterium,Vibrio parahaemolyticus,Staphylococcus aureus and others are used for detection among 49 samples which are 20% of various products samples with random selection.1 Salmonella,2 Staphylococcus aureus,1 Listeria monocytogenes bacterium and 1 Vibrio parahaemolyticus were detected out,with the detection rate of 10%.Conclusion： Compared to conventional bacterial culture,FQ-PCR is applicable to food-borne pathogens detection,which could increase detection rate and also reduce detection time.