实时荧光PCR在食源性疾病中的应用

目的:探讨实时荧光PCR快速检测在食物中毒中病原学基因诊断的应用。方法:采用实时荧光PCR检测技术对一起副溶血性弧菌引起的食物中毒患者的粪便和肛拭增菌液进行快速检测,同时采用传统的分离方法和细菌生化鉴定仪的生化鉴定,及日本生研株式会社生产的肠炎溶血性弧菌型别免疫血清(K型别)分型。结果:在这一起食物中毒的患者中,实时荧光PCR检测6份粪便和肛拭增菌液,其中,4份阳性,并从PCR阳性的增菌液中分离到4株副溶血性弧菌K6型。结论:实时荧光定量PCR技术特异性强,灵敏度高,能在采样后数小时内检测到病原菌的DNA,达到快速诊断的目的,在食物中毒的快速诊断上起重要作用。

Application of real - time quantitative PCR for rapid detection of pathogen in foods samples

Objective:To research the application of Real-Time Quantitative PCR(RQ-PCR) in rapidly detecting pathogenic bacteria genes causing food poisoning.Methods:We adopted the RQ-PCR to rapidly detect the feces and enrichment broth of anal swab which came from the victims of food poisoning caused by Vibrio parahaemolyticus.At the same time,we have identified the type of pathogenic Vibrio parahaemolyticus by the traditional bacterial separating technology and automatic bacterial biochemical identification system.We have identified the serotype by enteritis Vibrio parahaemolyticus diagnostic serum(type K) made by DENKA SEIKEN CO.,LTD.Results:Four of 6 samples from the feces and enrichment broth of anal swab were positive after detected by RQ-PCR.We also have separated 4 strains K6 type of Vibrio parahaemolyticus from the 4 positive sample.Conclusion:RQ-PCR is a sensitive and specific technique,it can detect the DNA of pathogenic bacteria in several hours after sampling.So the RQ-PCR can be used as a rapid detecting technique and will play an important part in the diagnosis of food poisoning.