福氏志贺菌4av和Yv血清型PCR鉴定方法的建立及应用

Development of PCR method for molecular serotyping of 4av and Yv of Shigella flexneri targeting on O-antigen modification genes

目的建立福氏志贺菌4av和Yv血清型PCR鉴定方法。方法根据福氏志贺菌4av和Yv血清型O抗结构,针对O抗合成基因wzx、IV型抗原决定基因gtrIV和MASF IV-1抗原决定基因opt,建立血清型4av和Yv的PCR鉴定方法。并应用该方法对126株福氏志贺菌临床分离株进行血清型检测。结果建立了一种福氏志贺菌4av和Yv血清型PCR鉴定方法,在一个反应中包括4对引物,Yv血清型PCR扩增为wzx及opt阳性;4av血清型PCR扩增为wzx、opt及gtrIV阳性。该方法可将4av和Yv血清型与目前已知的其他福氏志贺菌血清型完全区分。对126株不同血清型福氏志贺菌临床分离株的鉴定结果显示,该方法具有很好的特异性。结论本研究建立的福氏志贺菌4av和Yv血清型PCR鉴定方法,可以用于志贺菌检测和监测。

We developed a PCR assay for molecular serotyping of Shigella flexneri serotype 4av and Yv.Based on O-antigen structure of serotype 4av and Yv,primer pairs targeting on O-antigen synthesis gene wzx,the serotype 4-specific IV specific gene gtrIVand MASF IV-1specific gene opt were designed and included in one PCR assay to develop a molecular method for serotyping serotype 4av and Yv.Total of 126 S.flexneri clinical isolates were typed by this method and was compared with slide agglutination method.Results showed that a multiplex PCR assay was developed,containing four primer pairs in one single reaction could identify serotype Yv the target genes wzxand opt were positive and while 4av target genes,wzx,opt and gtrIV were all positive,being different from the others serotypes of S.flexneri.The PCR assay provides a rapid and specific method for S.flexneri serotype 4av and Yv identification,and could be well used in Shigelladetection and surveillance.