食品中空肠弯曲菌荧光重组酶聚合酶扩增检测方法的建立  

Development of a Detection Method for Food-borne Campylobacter jejuni Based on Fluorescence Probe-based Recombinase Polymerase Amplification

作　　者：兰全学 陈佳平 杨慧[1] 杜文旗 金玉娟[1] 甘莉萍[1] 刘渠[1] 刘小青[2] LAN Quan-xue;CHEN Jia-ping;YANG hui;DU Wen-qi;JIN Yu-juan;GAN Li-ping;LIU Qu;LIU Xiao-qing(Shenzhen Longgang District Center for Disease Control and Prevention.Shenzhen 518172,China;Shenzhen Academy of Metrology and Quality Inspection.Shenzhen 518109,China)

机构地区：[1]深圳市龙岗区疾病预防控制中心,广东深圳518172 [2]深圳市计量质量检测研究院,广东深圳518109

出　　处：《现代食品科技》2020年第5期304-309,共6页Modern Food Science and Technology

基　　金：深圳市科创委技术攻关项目(JSGG20160606144217004);广东省质量技术监督局项目(2018CZ20)。

摘　　要：本研究为了实现空肠弯曲菌的快速和便捷检测,建立了一种基于荧光重组酶聚合酶扩增技术(exo-RPA)快速检测空肠弯曲菌的方法。通过对空肠弯曲菌和对照菌株的exo-RPA检测来判断该方法的特异性。用梯度稀释的空肠弯曲菌作为模板进行检测来分析exo-RPA方法的灵敏度。通过对模拟污染样品检测来分析exo-RPA的应用效果。分别以exo-RPA和荧光PCR检测实际食品样品来分析二者的检测效果。空肠弯曲菌exo-RPA方法可特异检出空肠弯曲菌,检测灵敏度达到6.0×10^2CFU/m L。在模拟污染试验中,含2.5×10^1 CFU/m L空肠弯曲菌的增菌液在培养24 h后可以被exo-RPA检测出阳性信号。exo-RPA和荧光PCR对于40份样品的检测结果相同。本研究建立的空肠弯曲菌exo-RPA具有特异、灵敏和抗干扰性强的特点,方法操作简便快速,具有较好的应用前景。In order to achieve rapid and convenient detection of Campylobacter jejuni, a rapid detection method of Campylobacter jejuni based on fluorescence probe-based recombinase polymerase amplification(exo-RPA) was developed. The specificity of the method was determined by exo-PA detection of Campylobacter jejuni and control strains. The gradient-diluted Campylobacter jejuni was used as a template to analyze the sensitivity of the exo-RPA method. The application effect of the exo-RPA method was analyzed through detecting the simulated contaminated samples.The exo-RPA and fluorescence PCR were used to test real food samples respectively, in order to analyze their detection effects. The Campylobacter jejuni exo-RPA method can specifically detect Campylobacter jejuni with a detection sensitivity of 6.0×10^2 CFU/m L. In the simulated contamination tests, the enrichment broth medium containing 2.5×10^1 CFU/m L Campylobacter jejuni was tested positive after 24 hours of culture by the exo-RPA assay. The exo-RPA and fluorescence PCR methods led to the same detection results for the 40 samples. The Campylobacter jejuni exo-RPA assay developed in this study has high specificity, sensitivity and anti-interference ability. The method is simple, convenient and rapid, with a good application prospect.

关 键 词：重组酶聚合酶扩增技术 空肠弯曲菌 hipO基因 

分 类 号：O657.3[理学—分析化学] TS207.4[理学—化学]