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一种使用通用引物检测 11 种食源性致病菌的多重 PCR 检测方法。

A multiplex PCR assay with a common primer for the detection of eleven foodborne pathogens.

机构信息

Laboratory of Molecular Diagnostics, College of Pharmaceutical Sciences, Soochow Univ., Suzhou, Jiangsu, China.

Suzhou Center for Disease Control and Prevention, Suzhou, Jiangsu, China.

出版信息

J Food Sci. 2020 Mar;85(3):744-754. doi: 10.1111/1750-3841.15033. Epub 2020 Jan 30.

Abstract

Salmonella enterica, Listeria monocytogenes, Shigella flexneri, Escherichia coli O157:H7, Vibrio parahaemolyticus, Staphylococcus aureus, Vibrio cholerae, Clostridium botulinum type A, Bacillus cereus, Clostridium perfringens Alpha toxin, and Yersinia enterocolitica are 11 common foodborne pathogens. Traditional bacterial culture methods for detecting pathogens are time-consuming and labor-intensive. Multiplex PCR technology, which can detect multiple targets in a single tube, has been increasingly applied to microbial detection due to its high specificity, sensitivity, and fast response. This paper is to establish a multiplex PCR technology mediated by a common primer for the detection of these 11 common foodborne pathogens in order to achieve the goal of nondirectional screening for these 11 common foodborne pathogens. The specificity of the established CP-MPCR detection system was first verified by 100 clinical isolates. The sensitivity of the CP-MPCR detection system was then detected by using cultured bacteria preparations and has been confirmed with a high sensitivity of 10 to 10 CFU/mL, among them, the sensitivity of the CP-MPCR for Vibrio cholerae and S. flexneri can even achieve 10 CFU/mL. Sixty anal swab samples collected from Suzhou CDC and 16 enrichment cultured solutions of food samples collected from the Suzhou Food Inspection and Testing Center were tested using the CP-MPCR system. A total of 32 positive results were detected. PRACTICAL APPLICATION: Food poisoning incidents occur frequently around the world, mainly because of the contamination of food by pathogenic bacteria and serious harm to human health. The method provided in this study can detect 11 foodborne pathogens in food, which can effectively prevent the spread of pathogenic microorganisms. At the same time, for the food poisoning incident that has already occurred, this method can be used for diagnosis to find out the cause.

摘要

肠炎沙门氏菌、单增李斯特菌、福氏志贺菌、大肠杆菌 O157:H7、副溶血性弧菌、金黄色葡萄球菌、霍乱弧菌、A型肉毒梭菌、蜡样芽胞杆菌、产气荚膜梭菌α毒素和小肠结肠炎耶尔森菌是 11 种常见的食源性致病菌。传统的细菌培养方法检测病原体耗时耗力。由于其特异性高、灵敏度高、反应迅速,多重 PCR 技术已越来越多地应用于微生物检测,该技术可在单个管中检测多个靶标。本文旨在建立一种由通用引物介导的多重 PCR 技术,用于检测这 11 种常见食源性致病菌,以实现对这 11 种常见食源性致病菌的非定向筛选。首先通过 100 株临床分离株验证了所建立的 CP-MPCR 检测系统的特异性。然后通过培养细菌制剂检测 CP-MPCR 检测系统的灵敏度,已证实其灵敏度为 10 到 10 CFU/mL,其中 CP-MPCR 对霍乱弧菌和福氏志贺菌的灵敏度甚至可达 10 CFU/mL。使用 CP-MPCR 系统检测了来自苏州疾控中心的 60 份肛拭子样本和苏州食品检验检测中心采集的 16 份食品富集培养物。共检测到 32 个阳性结果。实际应用:世界各地经常发生食物中毒事件,主要是由于食物被致病菌污染,对人类健康造成严重危害。本研究提供的方法可以检测食品中的 11 种食源性致病菌,可以有效防止致病菌的传播。同时,对于已经发生的食物中毒事件,该方法可用于诊断,以找出病因。

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