高分辨率溶解曲线检测9种食源性致病菌方法的建立

Simultaneous Detection of Nine Foodborne Pathogenic Bacteria Using High-resolution Melting Analysis

摘要:

当今食源性疾病已成为一个全球关注的食品卫生问题，常见的细菌性食物中毒的病原菌有：致病性大肠杆菌（特别是出血性大肠杆菌O157:H7）、沙门氏菌、志贺氏菌、致病性弧菌（包括霍乱弧菌和副溶血弧菌）、金黄色葡萄球菌、单增李斯特菌、空肠弯曲菌以及阪崎克罗诺杆菌等。单独检测一种致病菌已无法满足现今社会对同时高效检测多种致病菌的要求，为建立一种同时对上述9种食品中常见致病菌的检测方法，本研究分别以上述9种菌的特异性基因为靶基因，创新性地结合一对通用引物，建立能同时检测多种食源性致病菌的多重HRM-real time PCR检测体系。结果表明该多重HRM-real time PCR检测体系通过两管PCR反应可对上述9种食品中常见致病菌进行有效的检测与区分，且具有良好的特异性和灵敏度。

Abstract:

Foodborne diseases have become a global food safety concern. Common bacterial pathogens involved in food poisoning include pathogenic Escherichia coli (especially enterohemorrhagic Escherichia coli O157:H7), Salmonella, Shigella, pathogenic Vibrio (including Vibrio cholerae and Vibrio parahaemolyticus), Staphylococcus aureus, Listeria monocytogenes, Campylobacter jejuni, and Cronobacter sakazakii. Species-specific detection for one type of pathogen cannot meet the requirements of modern society for multiple-pathogen detection. In order to establish a method that can simultaneously detect the above-mentioned nine pathogenic bacteria in food, a multiple high-resolution melting (HRM) real-time polymerase chain reaction (PCR) assay for the simultaneous detection of multiple foodborne pathogens was established, using the specific genes of the nine aforementioned bacteria as target genes and a novel pair of universal primers. The results showed that this multiple HRM-real time PCR detection system could effectively detect and distinguish these nine common pathogenic bacteria in food via a two-tube PCR reaction, with good specificity and sensitivity.