Submitted: 23 Jul 2014
Revised: 05 Nov 2014
Accepted: 14 Jan 2015
First published online: 06 Oct 2016
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Int J Enteric Pathog. 2015;3(2): e21796.
doi: 10.17795/ijep21796
  Abstract View: 464
  PDF Download: 422

Research Article

Molecular Detection and Speciation of Campylobacter Species in Children With Gastroenteritis Using Polymerase Chain Reaction in Bahonar Hospital of Karaj City

Naser Harzandi 1 * , Shima Jamshidi 1, Mehrouz Dezfulian 1, Alireza Bahonar 2, Amir Bakhtiari 1, Kamelia Banihashemi 1

1 Department of Microbiology, Karaj Branch, Islamic Azad University, Karaj, IR Iran
2 Department of Food Hygiene and Quality Control, Faculty of Veterinary Medicine, University of Tehran, Tehran, IR Iran
*Corresponding author: Naser Harzandi, Department of Microbiology, Karaj Branch, Islamic Azad University, Karaj, IR Iran. Tel: +98-9123481676, Fax: +98-2188283460, Email: naser.harzandi@kiau.ac.ir

Abstract

Background: Campylobacter spp. is recognized as one of the important bacterial agents of food borne diseases worldwide. C. jejuni and C. coli are the most frequently reported species causing bacterial gastrointestinal disorders in developed and some of the developing countries.

Objectives: The goals of this survey were to detect and to differentiate C. jejuni and C. coli species in stool specimens and to determine the frequency of Campylobacter gastroenteritis in children referred to Bahonar hospital in Karaj city using polymerase chain reaction (PCR).

Patients and Methods: One hundred sixty stool specimens were collected from neonates and children under 8 years old during August to October of 2009. PCR was optimized to amplify a 400 bp fragment of the cad F gene of Campylobacter genus in the clinical specimens, then multiplex PCR optimization was carried out using hipO and asp primers. C. jejuni RTCC 1097 and C. coli RTCC 1113 reference strains were used as positive controls and only the PCR positive specimens were examined by this method.

Results: The results revealed the amplification of a 400 bp DNA fragment and Campylobacter contamination in 11.3 % of the specimens. Of 18 PCR positive specimens examined by duplex PCR method, 4 (22.2%) were identified as C. jejuni, 2 (11.1%) as C. coli, 3 (16.6%) as mixed infection with both species and 9 (50% of the positive specimens) were identified as non coli – non jejuni Campyltobacter. The sensitivity of the single PCR method at the DNA level was determined to be 100 pg and the specificity of the method was determined using 6 other important bacterial agents of gastrointestinal diseases.

Conclusions: Although the findings of this survey confirmed the presence of C. jejuni and C. coli species in half of the positive specimens, the probable role of the other species of Campylobacter in children gastroenteritis was unexpected.

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