Department of Microbiology and Infectious Diseases, Faculté de Médecine et des Sciences de la Santé de l'Université de Sherbrooke, Sherbrooke, Québec, Canada.
PLoS One. 2011 Jan 24;6(1):e16167. doi: 10.1371/journal.pone.0016167.
Multi-locus sequence typing (MLST) has emerged as the state-of-the-art method for resolving bacterial population genetics but it is expensive and time consuming. We evaluated the potential of high resolution melting (HRM) to identify known MLST alleles of Campylobacter jejuni at reduced cost and time. Each MLST locus was amplified in two or three sub fragments, which were analyzed by HRM. The approach was investigated using 47 C. jejuni isolates, previously characterized by classical MLST, representing isolates from diverse environmental, animal and clinical sources and including the six most prevalent sequence types (ST) and the most frequent alleles. HRM was then applied to a validation set of 84 additional C. jejuni isolates from chickens; 92% of the alleles were resolved in 35 hours of laboratory time and the cost of reagents per isolate was $20 compared with $100 for sequence-based typing. HRM has the potential to complement sequence-based methods for resolving SNPs and to facilitate a wide range of genotyping studies.
多位点序列分型（MLST）已成为解析细菌群体遗传学的最新方法，但它既昂贵又耗时。我们评估了高分辨率熔解（HRM）在降低成本和时间的情况下识别空肠弯曲菌已知 MLST 等位基因的潜力。每个 MLST 基因座都被扩增为两个或三个亚片段，然后通过 HRM 进行分析。该方法使用 47 株空肠弯曲菌进行了研究，这些菌株先前通过经典 MLST 进行了表征，代表了来自不同环境、动物和临床来源的菌株，包括六个最常见的序列类型（ST）和最常见的等位基因。然后将 HRM 应用于来自鸡的 84 株空肠弯曲菌验证集；35 小时的实验室时间内解决了 92%的等位基因，每个分离株的试剂成本为 20 美元，而基于序列的分型为 100 美元。HRM 有可能补充基于序列的方法来解析 SNP，并促进广泛的基因分型研究。
