CAJ | 학술논문

目的建立基于O抗修饰基因的福氏志贺菌血清型PCR鉴定方法.方法根据福氏志贺菌O抗原合成及修饰特异基因设计引物,以常见的14种福氏志贺菌血清型(包括1a、1b、1c、2a、2b、3a、3b、4a、4b、5a、Y、X、Xv和F6)为标准菌株,建立福氏志贺菌血清型单重PCR鉴定方法；并以痢疾志贺菌、宋内志贺菌、鲍氏志贺菌和腹泻相关的其他菌属菌株验证特异性；应用该方法对106株福氏志贺菌临床分离株进行PCR血清分型.结果建立一种福氏志贺菌血清型单重PCR鉴定方法,通过8个PCR反应,能够鉴定目前已知的15种血清型中的14种(Xv除外).检测灵敏度在10 pg至1 ng DNA(20μl反应体系).对106株福氏志贺菌临床分离株的检测结果提示,PCR分型方法与玻片凝集法具有很高的一致性.结论本研究建立的福氏志贺菌血清型单重PCR鉴定方法具有特异性、灵敏性,可用于临床检测.
목적 건립기우O항수식기인적복씨지하균혈청형PCR감정방법.방법 근거복씨지하균O항원합성급수식특이기인설계인물,이상견적14충복씨지하균혈청형(포괄1a、1b、1c、2a、2b、3a、3b、4a、4b、5a、Y、X、Xv화F6)위표준균주,건립복씨지하균혈청형단중PCR감정방법；병이이질지하균、송내지하균、포씨지하균화복사상관적기타균속균주험증특이성；응용해방법대106주복씨지하균림상분리주진행PCR혈청분형.결과 건립일충복씨지하균혈청형단중PCR감정방법,통과8개PCR반응,능구감정목전이지적15충혈청형중적14충(Xv제외).검측령민도재10 pg지1 ng DNA(20μl반응체계).대106주복씨지하균림상분리주적검측결과제시,PCR분형방법여파편응집법구유흔고적일치성.결론 본연구건립적복씨지하균혈청형단중PCR감정방법구유특이성、령민성,가용우림상검측.
Objective To develop a singleplex PCR assay targeting O-antigen modification genes for molecular serotyping of Shigella (S.)flexneri.Methods Eight pairs of primer for O-antigen synthesis and modification genes of S.flexneriwere designed and used for developing an O-antigen modification gene-specific singleplex PCR assay to serotype 14 most common S.flexneri serotypes (1 a,1 b,1 c,2a,2b,3a,3b,4a,4b,5a,Y,X,Xv and F6).Bacterial pathogens which causing diarrheal disease were used for specificity detection.106 S.flexneri clinical isolates were serotyped by this method and compared with the slide agglutination method.Results An O-antigen modification,gene-specific singleplex PCR was developed.When six singleplex PCR reactions were performed,14 of the 15 recognized S.flexneri serotypes were identified,except for serotype Xv.The detection threshold ranged from 10 pg to 1 ng DNA in a 20 μ l reaction system.A high concordance between the singleplex PCR assay and slide agglutination were observed when 106 S.flexneri strains of various serotypes were analyzed with an exception that 1 serotype Y strain showed that it was carrying the additional defective gtr Ⅱ genes.Conclusion This method showed advantages over the traditional slide agglutination methods,and was promising when under application in the following situations as clinical diagnosis.