Evaluation of a Multiplex PCR Assay for the Identification of Campylobacter jejuni and Campylobacter coli

Authors
S. Shams 1
M. Ghorbanalizadgan 2
S. Haj Mahmmodi 3
A. Piccirillo 4
1 Cellular and Molecular Research Center, Qom University of Medical Sciences, Qom, Iran And Department of Medical Bacteriology, Faculty of Medical Sciences, Tarbiat Modares University, Tehran, Iran
2 Department of Bacteriology, Faculty of Medical Sciences, Baqiyatallah University, Tehran, Iran
3 Laboratory of Children's Medical Center. Tehran, Iran
4 5Department of Comparative Biomedicine and Food Science, University of Padua, Padova, Italy
Abstract
Background: Campylobacter jejuni and Campylobacter coli are identified as the major causes of acute gastroenteritis in humans. Because of the fastidious nature of Campylobacters, many clinical laboratories fail to routinely culture them. The detection of Campylobacter spp. using molecular-based techniques can be useful for diagnostic and epidemiological applications. This study aimed to developa multiplex PCR assay for the simultaneous detection of C. jejuni and C. coli strains from clinical specimens
Materials and Methods: During a 19-month period, stool samples were collected from 980 children admitted to a hospital in Tehran, Iran and then examined. The samples were cultured on both Brucella agar and Modified Charcoal-Cefoperazone-Deoxycholate agar (mCCDA) media at 42°C for 48 h. To confirm suspected bacteria, Gram staining and other biochemical tests were carried out. Finally, after extracting DNA from pure cultures using the boiling method, the multiplex PCR assay was performed.
Results: The multiplex PCR assay showed that Campylobacter spp. can be detected using 400 bp target product of cadF. It can also accurately distinguish between C. jejuni and C. coli species with different bands of 735 bp and 500 bp using hipO and asp genes, respectively
Conclusions: Results showed that the multiplex PCR assay can replace the biochemical assays for differentiating between C. jejuni and C. coli strains in a single-step PCR test.

Keywords

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