两种基于重组酶介导扩增技术的沙门氏菌检测方法比较

Comparison of Two Salmonella Detection Methods Based on Recombinase Aided Amplification Technology

该研究旨在利用重组酶介导扩增技术建立并比较两种食品中沙门氏菌快速检测方法。根据沙门氏菌菌毛蛋白fimY基因设计引物和探针,分别采用荧光法和侧向流试纸条法对扩增产物进行检测,构建重组酶介导等温核酸扩增方法,验证两种方法的特异性、灵敏度及对人工污染样品的检测效果。该研究构建的两种方法均在39℃下恒温运行,检测时间30~40 min,方法特异性良好,与大肠埃希氏菌、金黄色葡萄球菌等常见的其他食源性致病菌无交叉反应,侧向流试纸条法灵敏度可达到1 pg/μL,荧光法灵敏度为10 pg/μL,对人工污染样品的检测结果与传统分离培养法检测结果一致。因此该研究建立了两种快速、特异、灵敏的方法用于检测沙门氏菌,为食品中沙门氏菌污染的快速筛查提供一定的参考价值和数据支撑。

Two rapid detection methods for Salmonella in food were established and compared using recombinase-mediated amplification technology.The primers and probes were designed based on the fimY gene of Salmonella,and the amplified products were detected using the fluorescence method and lateral flow strip method,respectively.A recombinase-mediated isothermal nucleic acid amplification method was constructed to verify the specificity,sensitivity,and detection effect of the two methods on artificially contaminated samples.The two methods constructed in this study were operated at a constant temperature of 39℃,and the detection time was 30~40 min.These methods have good specificity and no cross-reaction with other common food-borne pathogens,such as Escherichia coli and Staphylococcus aureus.The sensitivity of the lateral flow test strip method reached 1 pg/μL,and the sensitivity of the fluorescence method was 10 pg/μL.The detection results of the artificially contaminated samples were consistent with the detection results of the traditional separation and culture method.Therefore,two rapid,specific and sensitive methods were established in this study for the detection of Salmonella,which provide a confirmed reference value and data to support the rapid screening of Salmonella contamination in food.