923 Multiplex PCR for Detection of Shiga-Like Toxin and Plasmid Antigen H Genes of Diarrheagenic Escherichia Coli

Abstract

Background and Aims Despite the fact that Shiga-like toxin E. coli has been identified as a major etiologic agent of children with diarrhea worldwide, few studies have been performed to evaluate the etiology of Shiga-like toxin-producing Escherichia coli (SLTEC) in children with diarrhea, in Iran. The aim of this study was to evaluate the etiology of Shiga-like toxin-producing Escherichia coli (SLTEC) in children with diarrhea, in Iran. Method A total of 300 stool specimens from children of 300 children with diarrhea were tested for the detection of E. coli, according to standard methods. Out of 300 specimens, 39 were identified as diarrheagenic E. coli, and subjected to multiplex polymerase chain reaction (MPCR) for detection of stx1/stx2, eae and ipaH genes. We designed a single multiplex polymerase chain reaction (MPCR) for the detection of target genes in diarrheagenic Escherichia coli. Results EPEC was the dominated strain (55.6%) among the tested isolates, followed by EHEC (25%) and EIEC (19.4%) strains. Conclusions Shiga-like toxin E. coli has been identified as a major etiologic agent of children with diarrhea in Iran. our method proved to be specific and rapid in detecting virulence genes from Shiga toxin-producing (stx1, stx2, and eae), enteropathogenic (eae), enteroinvasive (ipaH) Escherichia coli in stool samples, and able to simultaneously detect of diarrheagenic E. coli strains in a single reaction.