24重荧光real-time PCR技术在食物中毒快速检测中的应用

Application of 24-Plex Fluorescence Real-Time PCR Technology in Rapid Detection of Food Poisoning

目的:应用24重荧光real-time PCR检测技术快速筛检食物中毒病原菌,结合国家标准中的培养法探讨24重荧光real-time PCR检测技术符合性和应用价值。方法:采用高灵敏度、高特异性的24重荧光real-time PCR检测技术作为中毒病原菌的初筛方法,国标方法进行细菌分离培养,并对分离出的病原菌进行生化鉴定。结果:5份食物中毒患者肛拭子在增菌前检出4份霍乱弧菌核酸(非O1/非O139群,24重荧光real-time PCR),9份患者肛拭子在增菌后检出5株霍乱弧菌(非O1/非O139群,国标培养法),其中包含4份PCR技术初筛阳性样品,两个方法的符合率为80%。所有样本均未检出沙门氏菌、志贺菌、副溶血性弧菌、致泻性大肠埃希菌以及金黄色葡萄球菌。结论:应用24重荧光real-time PCR检测技术同时检测24种常见致病病原菌,能高效锁定中毒病原菌。将其与国标培养法相结合,对临床治疗和食物中毒快速处置能起到积极作用,值得应用和推广。

Objective:To use 24-plex fluorescence real-time PCR detection technology to rapidly screen food poisoning pathogens,and to explore the compliance and application value of 24-plex fluorescence real-time PCR detection technology combined with national standard culture method.Method:The 24-plex fluorescence real-time PCR detection technology with high sensitivity and specificity was used as the primary screening method for toxic pathogens,and the bacteria were isolated and cultured by national standard method,and the isolated pathogenic bacteria were biochemically identified.Result:Four samples of Vibrio cholerae nucleic acid(non O1/non O139 group,24-plex fluorescence real-time PCR)were detected from the anal swabs of 5 food poisoning patients before bacterial enrichment.Nine samples of patients detected five strains of Vibrio cholerae(non O1/non O139 group,national standard culture method)from their anal swabs after bacterial enrichment,including four samples that were initially screened positive by PCR technology.The compliance rate of the two methods was 80%.All samples did not detect Salmonella,Shigella,Vibrio parahaemolyticus,Escherichia coli,or Staphylococcus aureus.Conclusion:The 24-plex fluorescence real-time PCR detection technology was used to detect 24 common pathogenic bacteria at the same time,which could effectively lock the poisoning pathogens.Combining it with national standard culture method can play a positive role in clinical treatment and rapid treatment of food poisoning,which is worthy of application and promotion.