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