Department of Medical Microbiology, School of Medicine, University of Manchester, Clinical Sciences Building, Manchester Royal Infirmary, Oxford Road, Manchester, M13 9WL, United Kingdom.
J Microbiol Methods. 2010 Apr;81(1):11-6. doi: 10.1016/j.mimet.2010.01.013. Epub 2010 Jan 28.
The accurate sub-typing of Salmonella enterica isolates is essential for epidemiological investigations and surveillance of Salmonella infections. Salmonella isolates are currently identified using the Kauffman-White serotyping scheme. Multilocus sequence typing (MLST) schemes have been developed for the major bacterial pathogens including Salmonella and have assisted in understanding the molecular epidemiology and population biology of these organisms. Recently, the DiversiLab rep-PCR system has been developed using micro-fluidic chips to provide standardized, semi-automated fingerprinting for pathogens including S. enterica. In the current study, 71 isolates of S. enterica, representing 21 serovars, were analyzed using MLST and the DiversiLab rep-PCR system. MLST was able to identify 31 sequence types (STs), while the DiversiLab system revealed 38 DiversiLab types (DTs). The rep-PCR distinguished isolates of different serovars and showed greater discriminatory power (0.95) than MLST typing (0.89). Rep-PCR exhibited 92% concordance with MLST and 90% with serotyping, while the concordance level of MLST typing with serotyping was 96%, representing a strong association. Comparison of rep-PCR profiles with those held in an online library database led to the accurate prediction of serovar in 63% of cases and resulted in inaccurate predictions for 10% of profiles. MLST and the rep-PCR system may provide useful additional informative techniques for the molecular identification of S. enterica. We conclude that the DiversiLab rep-PCR system may provide a rapid (less than 4h) and standardized method for sub-typing isolates of S. enterica.
准确鉴定沙门氏菌分离株对于沙门氏菌感染的流行病学调查和监测至关重要。沙门氏菌分离株目前采用考夫曼-怀特血清分型方案进行鉴定。多位点序列分型(MLST)方案已针对包括沙门氏菌在内的主要细菌病原体开发,并有助于了解这些生物体的分子流行病学和种群生物学。最近,DiversiLab 重复序列扩增(rep-PCR)系统已使用微流控芯片开发,为包括 S. enterica 在内的病原体提供标准化、半自动化的指纹图谱分析。在本研究中,对 71 株代表 21 种血清型的 S. enterica 分离株进行了 MLST 和 DiversiLab rep-PCR 系统分析。MLST 能够鉴定出 31 种序列型(ST),而 DiversiLab 系统则显示出 38 种 DiversiLab 型(DT)。rep-PCR 能够区分不同血清型的分离株,且具有比 MLST 分型更高的分辨力(0.95)。rep-PCR 与 MLST 分型的一致性为 92%,与血清分型的一致性为 90%,而 MLST 分型与血清分型的一致性水平为 96%,表明存在很强的关联。将 rep-PCR 图谱与在线文库数据库中的图谱进行比较,能够准确预测 63%的血清型,而对 10%的图谱则预测不准确。rep-PCR 和 MLST 系统可能为沙门氏菌的分子鉴定提供有用的补充信息技术。我们得出结论,DiversiLab rep-PCR 系统可能为沙门氏菌分离株的快速(少于 4 小时)和标准化分型提供一种方法。