Login | Request Account (DAF staff only)

Campylobacter jejuni and Campylobacter coli Genotyping by High-Resolution Melting Analysis of a flaA Fragment.

Share this record

Add to FacebookAdd to LinkedinAdd to XAdd to WechatAdd to Microsoft_teamsAdd to WhatsappAdd to Any

Export this record

View Altmetrics

Merchant-Patel, S., Blackall, P. J., Templeton, J. M., Price, E.P., Tong, S.Y.C., Huygens, F. and Giffard, P.M. (2010) Campylobacter jejuni and Campylobacter coli Genotyping by High-Resolution Melting Analysis of a flaA Fragment. Applied and Environmental Microbiology, 76 (2). pp. 493-499.

Full text not currently attached. Access may be available via the Publisher's website or OpenAccess link.

Article Link: http://dx.doi.org/10.1128/AEM.01164-09

Publisher URL: http://www.asm.org

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 this primer set was used for the remainder of the study. Eighty-seven C. jejuni and 15 C. coli isolates were analyzed by flaA HRM and also partial flaA sequencing. There were 47 flaA sequence variants, and all were resolved by HRM analysis. The isolates used had previously also been genotyped using single-nucleotide polymorphisms (SNPs), binary markers, CRISPR HRM, and flaA RFLP.flaA HRM analysis provided resolving power multiplicative to the SNPs, binary markers, and CRISPR HRM and largely concordant with the flaA RFLP. It was concluded that HRM analysis is a promising approach to genotyping based on highly variable genes.

Item Type:Article
Business groups:Animal Science
Keywords:Single-nucleotide polymorphisms; flagellin genes; recombination; region; resistance; markers; HRM.
Subjects:Science > Biology > Genetics
Science > Microbiology > Bacteria
Live Archive:28 Jun 2010 04:52
Last Modified:04 Nov 2022 03:28

Repository Staff Only: item control page