Best E L, Fox A J, Owen R J, Cheesbrough J, Bolton F J
Laboratory of Enteric Pathogens, Centre for Infections, Health Protection Agency, London, UK.
Epidemiol Infect. 2007 Jul;135(5):839-46. doi: 10.1017/S0950268806007461. Epub 2006 Nov 17.
Specimens of human faeces were tested by a rapid strategy for detection of Campylobacter jejuni lineages by the presence of specific single nucleotide polymorphisms (SNPs) based on the C. jejuni multi locus sequence typing (MLST) scheme. This strategy was derived from analysis of the MLST databases to identify clonal complex specific SNPs followed by the design of real-time PCR assays to enable identification of six major C. jejuni clonal complexes associated with cases of human infection. The objective was to use the MLST SNP-based assays for the direct detection of C. jejuni by clonal complex from specimens of human faeces, and then confirm the accuracy of the clonal complex designation from the SNP-based assays by performing MLST on the cultured faecal material, this targeted at determining the validity of direct molecular specimen identification. Results showed it was possible to identify 38% of the isolates to one of the six major MLST clonal complexes using a rapid DNA extraction method directly from faeces in under 3 h. This method provides a novel strategy for the use of real-time PCR for detection and characterization beyond species level, supplying real-time epidemiological data, which is comparable with MLST results.
通过基于空肠弯曲菌多位点序列分型(MLST)方案,利用特定单核苷酸多态性(SNP)的存在,采用快速策略对人类粪便样本进行空肠弯曲菌谱系检测。该策略源自对MLST数据库的分析,以识别克隆复合体特异性SNP,随后设计实时PCR检测方法,以鉴定与人类感染病例相关的六种主要空肠弯曲菌克隆复合体。目的是使用基于MLST SNP的检测方法直接从人类粪便样本中按克隆复合体检测空肠弯曲菌,然后通过对培养的粪便材料进行MLST来确认基于SNP检测方法的克隆复合体指定的准确性,这旨在确定直接分子样本鉴定的有效性。结果表明,使用快速DNA提取方法,在3小时内直接从粪便中能够将38%的分离株鉴定为六种主要MLST克隆复合体之一。该方法为使用实时PCR进行物种水平以上的检测和表征提供了一种新策略,提供与MLST结果相当的实时流行病学数据。