Molecular Detection and Characterization of Diarrheagenic Escherichia coli from Clinical isolates in Khartoum State

Abstract

Diarrheagenic Escherichia coli is one of the most important etiologic agents of several infections, and represent a major public health problem in developing countries like Sudan. The present study was undertaken to detect the virulence factors of diarrheagenic E. Coli strains using multiplex PCR in isolated samples of urine, high vaginal swab and wound swab. Fifty clinical isolate were collected from Omdurman military hospital and police hospital in Khartoum State; they were from 21(42%) males and 29(58%) females. Then they cultured on MacConkey agar and EMB media. Then biochemical tests were done, including indole test, citrate, Kilgelar iron agar and urease test. The sensitivity test was performed using Disc diffusion method; the following antibiotic discs were used (Ceftriaxone, Meropenem, Imipenem, Amikacin, Ceftazidime, Ciprofloxacin Gentamicin and Framycetin). Most of the isolated organisms were resistant to these antibiotics. DNA was extracted by boiling method. Multiplex PCR techniques was used to amplify four genes of diarrheagenicE.coli, the frequency of Enteroaggregative E.coli (aggRgene) was found in 8 (16) % in urine isolate , and negative in other isolates, Enterohermorrhagic E.coli (stxgene)was2(4%) in urine isolates and negative in other isolates,Enteroinvasive E.coli (ipaH gene) was (8) 16 % in urine isolate, and 2 (4%) in wound swab and 1(2%) in High vaginal swab. Enteropathogenic E.coli(eae gene) was negative in all isolates. From this study we concluded that, the presence of diarrheagenic E.coliin other samples than stools may indicate the source of infection and the predominant strain is Enteroinvasive E.coli .