CAJ | 학술논문

目的评价REP-PCR基因分型技术在临床光滑假丝酵母菌株分型中的应用价值。方法收集2009-2010年上海瑞金医院、上海仁济医院、上海华山医院、安徽医科大学附属医院、深圳人民医院38株光滑假丝酵母菌。采用MLST法扩增光滑假丝酵母菌的6个管家基因内片段，扩增片段测序后与数据库数据比对得出等位基因谱，从而得到相应的序列型（sequence type，ST）。采用REP-PCR法设计Ca21、Ca22、Com21引物扩增38株光滑假丝酵母菌重复序列，通过电泳比较分析，获得REP-PCR型。比较两种分型方法的结果和效率。结果以Ca22-Com21为引物的REP-PCR分型效果最好。REP-PCR和MLST分型结果相同，38株光滑假丝酵母菌共产生5种REP-PCR型，分别为A、B、C、D、E型，对应于MLST的ST7、ST3、ST19、ST45和新型。REP-PCR比MLST耗时短。结论REP-PCR方法简单快速，且分辨率与MLST技术相同，可作为实验室大量菌株分型的首选方法。
목적 평개REP-PCR기인분형기술재림상광활가사효모균주분형중적응용개치。방법수집2009-2010년상해서금의원、상해인제의원、상해화산의원、안휘의과대학부속의원、심수인민의원38주광활가사효모균。채용MLST법확증광활가사효모균적6개관가기인내편단，확증편단측서후여수거고수거비대득출등위기인보，종이득도상응적서렬형（sequence type，ST）。채용REP-PCR법설계Ca21、Ca22、Com21인물확증38주광활가사효모균중복서렬，통과전영비교분석，획득REP-PCR형。비교량충분형방법적결과화효솔。결과 이Ca22-Com21위인물적REP-PCR분형효과최호。REP-PCR화MLST분형결과상동，38주광활가사효모균공산생5충REP-PCR형，분별위A、B、C、D、E형，대응우MLST적ST7、ST3、ST19、ST45화신형。REP-PCR비MLST모시단。결론REP-PCR방법간단쾌속，차분변솔여MLST기술상동，가작위실험실대량균주분형적수선방법。
Objective To assess the application value of REP-PCR in genotyping of candida glabrata strains in clinical pratice. Methods From 2009 to 2010, thirty-eight candida glabrata strains were isolated from Shanghai Ruijin Hospitals, Shanghai Renji Hospital, Shanghai Huashan Hospital, Anhui Medical University Hospital, Shenzhen People's Hospital. Six loci in housekeeping genes (FKS, LEU2,NMT1, TRP1, UGP1 and URA3 ) were amplified and sequenced. The sequences were compared with the MIST database and allele profile and sequence type (ST) were obtained. With primers Ca21, Ca22 and Com21 used to amplify the adjacent variable gene regions, the amplicons were analyzed through electrophoresis to generate different REP-PCR types. Finally, the results of these two genotyping methods were compared. Results For REP-PCR, Ca22-Com21 has the best genotyping effect. REP-PCR and MLST have the same genotyping results. Five REP-PCR types were found in 38 candida glabrsta isolates. Type A,B, C, D and E strains from REP-PCR were genotyped as ST 7, 3, 19, 45 and new type respectively byMIST. REP-PCR saves time compared with MIST. Conclusions REP-PCR offers a simple and rapid method for molecular typing, with similar discriminatory power with MIST. Therefore, REP-PCR can be the preferred choice in laboratory, especially for a large number of isolates.