ArticleApplied and environmental microbiology2010
Campylobacter jejuni and Campylobacter coli genotyping by high-resolution melting analysis of a flaA fragment.
Article in Applied and environmental microbiology, 2010. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 13 papers.
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13 citing papers in PubMed.
- Investigation of Campylobacter colonization in three Australian commercial free-range broiler farms.Poultry science · 2021Article
- Identification and differentiation ofVeterinary world · 2020Article
- Genotyping of Campylobacter jejuni Isolates from Poultry by Clustered Regularly Interspaced Short Palindromic Repeats (CRISPR).Current microbiology · 2020Article
- High resolution melting analysis of the 18S rRNA gene for the rapid diagnosis of bovine babesiosis.Parasites & vectors · 2019Article
- Amino acid permease 3 (aap3) coding sequence as a target for Leishmania identification and diagnosis of leishmaniases using high resolution melting analysis.Parasites & vectors · 2018Article
- Detection and discrimination of two Brucella species by multiplex real-time PCR and high-resolution melt analysis curve from human blood and comparison of results using RFLP.Iranian journal of basic medical sciences · 2015Article
- Differentiation of Campylobacter jejuni and Campylobacter coli Using Multiplex-PCR and High Resolution Melt Curve Analysis.PloS one · 2015Article
- Pyrazinamide susceptibility testing of Mycobacterium tuberculosis by high resolution melt analysis.Tuberculosis (Edinburgh, Scotland) · 2014Article
- Application of HRM assays with EvaGreen dye for genotyping Giardia duodenalis zoonotic assemblages.Parasitology research · 2012Article
- Microbiological applications of high-resolution melting analysis.Journal of clinical microbiology · 2012Review
- Article
- Presence and characterization of Campylobacter jejuni in organically raised chickens in Quebec.Canadian journal of veterinary research = Revue canadienne de recherche veterinaire · 2011Article
- High-resolution melting system to perform multilocus sequence typing of Campylobacter jejuni.PloS one · 2011Article
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7 authors.
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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.
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