Hookey J V, Arnold C
Molecular Biology Unit, Sexually Transmitted and Blood Borne Virus Laboratory, Central Public Health Laboratory, 61 Colindale Avenue, London NW9 5HT.
J Med Microbiol. 2001 Nov;50(11):991-995. doi: 10.1099/0022-1317-50-11-991.
Five National Collection of Type Culture (NCTC) strains and 14 isolates of Neisseria meningitidis, representing 13 outbreak isolates from within the UK, were examined by multilocus sequence typing (MLST) for seven house-keeping genes. The results were compared with those of fluorescent amplified fragment-length polymorphism (FAFLP) analysis. Phylogenetic inferences were made from 3284-nucleotide lengths of sequence for the 19 isolates, by distance and parsimony methods. Two clusters of isolates were delineated. The larger, comprising eight isolates--S1, S3, Ironville, P9, ET-37 (M99-241951), P7, P10 and P60--shared 100-99.2% similarity and varied in only 40 nucleotides (approximately 1.22% variation) from the consensus sequence alignment. This cluster could be equated to the ET-37 complex because it had allelic signatures identical to MLST sequence types 11 and 50. These eight isolates were also assigned to one group by FAFLP. The reference ET-5 complex isolate 'ET-5 (NG144/82)' and an isolate (X9) from an outbreak in the north of England were also grouped together by MLST. They shared 99.2% similarity and differed within the aroE and fumC genes by 4 and 17 nucleotides, respectively. Their MLST sequence types were 32 and 661 and, therefore, these two isolates could be equated to the ET-5 complex. They also grouped together by FAFLP. A comparison of the resources required to apply MLST to the 19 isolates examined with those needed to characterise them by FAFLP indicated that FAFLP (a fragment-based genotyping method) is more cost-effective than the partial sequencing approach, MLST.
采用多位点序列分型(MLST)方法,对5株保藏于英国国家典型菌种保藏中心(NCTC)的菌株以及14株脑膜炎奈瑟菌分离株进行了7个管家基因的检测。这些分离株代表了来自英国境内的13株暴发菌株。研究结果与荧光扩增片段长度多态性(FAFLP)分析结果进行了比较。通过距离法和简约法,基于19株分离株3284个核苷酸长度的序列进行了系统发育推断。结果划分出两个分离株簇。较大的一个簇包含8株分离株——S1、S3、Ironville、P9、ET - 37(M99 - 241951)、P7、P10和P60,它们之间的相似性为100% - 99.2%,与共有序列比对仅相差40个核苷酸(约1.22%的变异)。该簇等同于ET - 37复合体,因为其等位基因特征与MLST序列类型11和50相同。这8株分离株通过FAFLP也被归为一组。参考ET - 5复合体分离株“ET - 5(NG144/82)”以及来自英格兰北部一次暴发的一株分离株(X9)通过MLST也被归为一组。它们的相似性为99.2%,在aroE和fumC基因内分别相差4个和17个核苷酸。它们的MLST序列类型分别为32和661,因此这两株分离株等同于ET - 5复合体。它们通过FAFLP也被归为一组。将对19株分离株应用MLST所需的资源与通过FAFLP对其进行分型所需的资源进行比较,结果表明FAFLP(一种基于片段的基因分型方法)比部分测序方法MLST更具成本效益。