Differentiation of Campylobacter jejuni and Campylobacter coli using multiplex-PCR and high resolution melt curve analysis
Banowary, Banya, Dang, Van Tuan, Sarker, Subir, Connolly, Joanne H., Chenu, Jeremy, Groves, Peter, Ayton, Michelle, Raidal, Shane, Devi, Aruna, Vanniasinkam, Thiru, and Ghorashi, Seyed A. (2015) Differentiation of Campylobacter jejuni and Campylobacter coli using multiplex-PCR and high resolution melt curve analysis. PLoS ONE, 10 (9). e0138808.
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Abstract
Campylobacter spp. are important causes of bacterial gastroenteritis in humans in developed countries. Among Campylobacter spp. Campylobacter jejuni (C. jejuni) and C. coli are the most common causes of human infection. In this study, a multiplex PCR (mPCR) and high resolution melt (HRM) curve analysis were optimized for simultaneous detection and differentiation of C. jejuni and C. coli isolates. A segment of the hippuricase gene (hipO) of C. jejuni and putative aspartokinase (asp) gene of C. coli were amplified from 26 Campylobacter isolates and amplicons were subjected to HRM curve analysis. The mPCR-HRM was able to differentiate between C. jejuni and C. coli species. All DNA amplicons generated by mPCR were sequenced. Analysis of the nucleotide sequences from each isolate revealed that the HRM curves were correlated with the nucleotide sequences of the amplicons. Minor variation in melting point temperatures of C. coli or C. jejuni isolates was also observed and enabled some intraspecies differentiation between C. coli and/or C. jejuni isolates. The potential of PCR-HRM curve analysis for the detection and speciation of Campylobacter in additional human clinical specimens and chicken swab samples was also confirmed. The sensitivity and specificity of the test were found to be 100% and 92%, respectively. The results indicated that mPCR followed by HRM curve analysis provides a rapid (8 hours) technique for differentiation between C. jejuni and C. coli isolates.
| Item ID: | 79871 |
|---|---|
| Item Type: | Article (Research - C1) |
| ISSN: | 1932-6203 |
| Copyright Information: | Copyright: © 2015 Banowary et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited |
| Date Deposited: | 14 Aug 2023 23:07 |
| FoR Codes: | 31 BIOLOGICAL SCIENCES > 3107 Microbiology > 310701 Bacteriology @ 100% |
| SEO Codes: | 28 EXPANDING KNOWLEDGE > 2801 Expanding knowledge > 280101 Expanding knowledge in the agricultural, food and veterinary sciences @ 50% 28 EXPANDING KNOWLEDGE > 2801 Expanding knowledge > 280103 Expanding knowledge in the biomedical and clinical sciences @ 50% |
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