实时荧光PCR检测志贺菌特异基因ipaH

Detection of Shigella with a real-time PCR assay targeting ipaH gene

目的:建立一种灵敏、快速检测志贺菌的实时荧光PCR方法,研究志贺菌在腹泻病人中的感染情况。方法:针对志贺菌特异性基因ipaH设计引物、探针,应用培养分离的志贺菌进行验证,同时检测360例临床标本并与培养法进行对照。结果:应用实时荧光PCR方法,10株志贺菌检测结果均为阳性,10株非志贺菌结果均为阴性;检测灵敏度达到4 cfu/test。360例临床标本中实时荧光PCR方法检测出29例阳性,培养法检测出12例阳性。结论:实时荧光PCR法检测志贺菌具有灵敏度高、快速方便的特点,可用于检测腹泻病人志贺菌感染。

Objective：To develop a specific and sensitive real -time PCR method for the detection of Shigella. Methods. Lyses of bacterial pellets from the samples were analyzed with real - time PCR based on TaqMan technology, The PCR assay targeted the ipaH gene specific for Shigella spp. Previous identified strains and 360 clinical samples were tested with real - time PCR and culture method. Results.The PCR test correctly identified all the 10 Shigella strains. No false negative result was found in 10 non - Shigella strains. The lower detection limit could be down to 5 ~ 102 cfu/ml （4 cfu per reaction）. Real - time PCR detected 29 positive results in 360 clinical samples, while culture identified 12 positive samples. Conclusion： The real -time TaqMan assay can be a sensitive and simple alternative method for identification of Shigella.