实时荧光聚合酶链式反应技术与国标方法检测致病菌的应用与研究

Application and research of real-time polymerase chain reaction and national standard method for detection of pathogenic bacteria

目的比较实时荧光聚合酶链式反应技术(real-time polymerase chain reaction,RT-PCR)与国标方法在食品致病菌检测中的异同。方法应用RT-PCR法及国标GB 4789系列对采集的60件畜产品、禽产品、水产品和乳及乳制品中的沙门氏菌、金黄色葡萄球菌、单核细胞增生李斯特氏菌以及副溶血性弧菌同时进行检测。结果除了金黄色葡萄球菌检测结果均为阴性外,其他3种致病菌2种方法都有检出,但RT-PCR方法的阳性率均高于国标方法。对于沙门氏菌,国标方法阳性率为1.67%,RT-PCR方法阳性率3.33%;单核细胞增生李斯特氏菌国标方法阳性率为7.50%,RT-PCR方法阳性率为15.00%;副溶血性弧菌国标方法阳性率为8.33%,RT-PCR方法阳性率为11.67%。结论在本实验条件下,RT-PCR技术的阳性检测结果多于国标GB4789系列,并且结果可完全覆盖国标GB4789。各企业实验室甚至政府主导的食品风险监测项目可根据产品特点,合理应用RT-PCR技术以减少人员工作量,方便产品放行并提高工作效率。

Objective To compare the similarities and differences between real-time polymerase chain reaction(RT-PCR)and national standard methods in the detection of food pathogens.Methods Salmonella,Staphylococcus aureus,Listeria monocytogenes and Vibrio parahaemolyticus in 60 livestock products,poultry products,aquatic products and dairy and dairy products were detected simultaneously by RT-PCR and GB 4789 series.Results Except for the negative results of Staphylococcus aureus,the other three pathogenic bacteria were detected by two methods,but the positive rate of RT-PCR method was higher than that of national standard method.For Salmonella,the positive rates of GB method and RT-PCR method were 1.67%and 3.33%,respectively.For Listeria monocytogenes,the positive rates of GB method and RT-PCR method were 7.50%and 15.00%,respectively.For Vibrio parahaemolyticus,the positive rates of GB method and RT-PCR method were 8.33%and 11.67%,respectively.Conclusion Under this experimental condition,RT-PCR technology has more positive detection results than the national standard GB 4789 series,and the results can completely cover the national standard GB 4789.Enterprise laboratories and even government-led food risk monitoring projects can reasonably apply RT-PCR technology to reduce personnel workload,facilitate product release and improve work efficiency based on product characteristics.