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

Comparative Study of Campylobacter spp. Isolated from Children With Gastroenteritis in Bahonar Hospital, Karaj, Using PCR and RFLP

“…Digestion of PCR products by AluI and TasI endonucleases yielded known electrophoretic patterns (the RFLP method confirms our PCR results) (15,16). …”

“…In this study PCR using genus-specific primers (cadf and 23S rRNA) was performed to determine the frequency of Campylobacter contamination in the cases of children with gastroenteritis in Bahonar hospital, Karaj, Iran, and PCR using species-specific primers (hipo for C. jejuni and asp for C. coli) and RFLP were carried out to differentiate the interfering species. The results of PCR using both genusspecific primers (cadf and 23S rRNA) revealed Campylobacter contamination in 20% of tested stool samples, which is relatively high rate compared to the results of most other published surveys, i.e., those performed by Bessede et al (15). The higher frequency of Campylobacter gastroenteritis could be due to increased consumption of undercooked poultry products in recent years as well as the high sensitivity of molecular techniques in comparison with culture-based methods.…”

“…Cadf, hipo, and asp genes have all been the targets of primers used in much independent research. Although these genes are reported to be highly conserved among isolates from food and clinical samples (9), according to the results of recent molecular surveys on Campylobacters (15,16) variation in the sequence of hipo and asp primer binding sites in Iranian isolates is probable, and the aim of this study was to test this hypothesis. Comparing the results of PCR and RFLP using AluI RE in the speciation of positive samples (Table 3) confirmed the above-mentioned hypothesis.…”

Multiplex PCR Assay for Identifi cation and Differentiation of Campylobacter jejuni and Campylobacter coli Isolates

“…C. jejuni has the gene hippuricase -hipO, but this gene might not be expressed, and the reaction in the phenotypic differentiation may be false positive. 19,20 The multiplex PCR assay was initially developed and optimized using two Campylobacter reference strains (C. jejuni ATCC 33560 and C. coli Denmark WHO 814). To evaluate the effectiveness of the multiplex PCR assay, all Campylobacter isolates previously identifi ed by conventional biochemical-based identifi cation were tested.…”

“…The strain then may be inaccurately identifi ed as C. coli. 19,20 This method will be further optimized to specifically identify C. jejuni and C. coli directly from feces which will reduce the time and cost of the diagnostic process in Bulgaria where the high incidence of gastrointestinal infections is still an important clinical and socio-economic problem.…”