Development and Evaluation of a Multiplex PCR for the Detection of Campylobacter concisus and Other Campylobacter spp. from Gastroenteritis Cases

Mohsina Huq, Gena Gonis, Taghrid Istivan

Bacteriology Laboratory, Microbiology, Department of Laboratory Services, The Royal Children's Hospital, Melbourne, Australia.
Biotechnology and Environmental Biology, School of Applied Sciences, RMIT University, Melbourne, Australia.

DOI: 10.4236/ojmm.2014.41005                

Abstract

We developed and evaluated a multiplex PCR (m-PCR) for application in routine diagnostic laboratories to detect Campylobacter spp. in stool samples including C. concisus, C. jejuni, and C. coli. When this m-PCR was applied on spiked faecal samples, C. concisus, C. jejuni, and C. coli were specifically identified at 105 cells/gm of faeces. To compare the sensitivity of the m-PCR with conventional culture techniques, the same spiked stool samples were cultured on an antibiotic free Columbia blood agar using the filtration technique. The detection limit of conventional culture method was 105 cells/gm of stool for C. concisus and 106 cells/gm of stool for C. jejuni and C. coli. The m-PCR was applied to test 127 faecal samples from children with gastroenteritis and the results were compared with the conventional bacterial cultures data. By this m-PCR technique, C. jejuni was detected in 7 samples, C. coli in 2 samples, and C. concisus in 7 samples. However, the conventional culture results for these samples were 6 for C. jejuni, 2 for C. coli and only one sample was positive for C. concisus. In total, 19 samples were positive for Campylobacter spp. by m-PCR while only 9 samples were positive for Campylobacter spp. by culture. In conclusion, m-PCR is more sensitive than the culture technique to detect C. concisus and other fastidious campylobacters in faeces.

Keywords

Campylobacter concisus; Campylobacter spp.; Gastroenteritis; Faecal Spiking; Multiplex PCR

Conflicts of Interest

The authors declare no conflicts of interest.

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