Simultaneous Molecular Detection of Common Bacterial Enteropathogens in Children with Diarrhea by Multiplex-PCR Assay

Background: Diarrhea remains a major threat to children in low- and middle-income countries, which is the second cause of death among children in the world. The aim of the present study was to develop and evaluate a multiplex-PCR assay for direct detection of common bacterial enteropathogens in fecal specimens. Methods: One hundred and three stool specimens were collected from children under 5 years of age with gastroenteritis during a six-month period in Ilam, Iran. The multiplex PCR assay simultaneously detected Shigella spp., Campylobacter jejuni, Enteropathogenic Escherichia coli (EPEC), Enterotoxigenic Escherichia coli (ETEC), and Salmonella enterica in stool samples. Results: Our results demonstrated that the prevalence of Shigella spp. Campylobacter jejuni, EPEC, ETEC, and Salmonella enterica were 21.35%, 10.67%, 1.94%, 0.97% and 0%, respectively. In addition, Shigella spp. with Campylobacter jejuni and EPEC with Campylobacter jejuni coinfection were observed in sample 11 (10.67%). The analytical sensitivity of the multiplex PCR assay was estimated to be 0.01 ng/mu L of genomic DNA from culture. The analytical specificity was determined to be 100% by using common and standard enteropathogenic bacterial strains. Conclusions: The molecular method developed in the study was rapid, sensitive, and specific for detection of common bacterial enteropathogens.