Campylobacter jejuni and Campylobacter coli genotyping by high-resolution melting analysis of a flaA fragment.

ABSTRACT: The highly variable flagellin-encoding flaA gene has long been used for genotyping Campylobacter jejuni and Campylobacter coli. High-resolution melting (HRM) analysis is emerging as an efficient and robust method for discriminating DNA sequence variants. The objective of this study was to apply HRM analysis to flaA-based genotyping. The initial aim was to identify a suitable flaA fragment. It was found that the PCR primers commonly used to amplify the flaA short variable repeat (SVR) yielded a mixed PCR product unsuitable for HRM analysis. However, a PCR primer set composed of the upstream primer used to amplify the fragment used for flaA restriction fragment length polymorphism (RFLP) analysis and the downstream primer used for flaA SVR amplification generated a very pure PCR product, and

SUBMITTER: Merchant-Patel S 

PROVIDER: S-EPMC2805211 | biostudies-literature | 2010 Jan

REPOSITORIES: biostudies-literature