Genotypic detection of the Virulence Factors of Uropathogenic Escherichia coli Isolated from Diarrheic and Urinary Tract Infected Patients in Khartoum State

Abstract

The aim of this study was to determine the virulence factor of UropathogenicE.coli isolated from diarrheic and urinary tract infected patients in Khartoum State by multiplex PCR assay. A total of 100 clinical specimens(50 urine, 50 diarrhea) were collected in this study. Urine samples were culture on CLED agar, while diarrhea samples were culture on MacConky agar,identification scheme was done by conventional method. Modified Kirby-bauer method was performed using the following antibiotic discs; Gentamicin,Amikacin, Ciprofloxacin and Co-trimoxazole.Fifty five percent of samples were found sensitive to Gentamicin, 96% were sensitive to Amikacin, 57% weresensitive to Ciprofloxacin and 63% were sensitive to Co-trimoxazole. Boiling method was adopted for DNA extraction.Finally Multiplex PCR was done for thedetection of E. colivirulent genes(pap, fim, sfa, aer and hly). Most study population were females 57(57%); 42 of them suffering from UTIs and 15 suffering from diarrhea, while males were 43(43%); 8 of them were suffering from UTIs and 35 of them were suffering from diarrhea. Among enrolled subjects, 82 were positive for one or more Uropathogenic E. coli virulent genes. While18 isolates were negative for all genes. The results of multiplex PCR searching for different virulent factors revealed the following:Thirty tow (n=32) diarrheal samples appear asaer gene positive while the remaining fourteen (n=14) urine samples appear as aer gene positive. Thirty three (n=33) urine samples appear as fim gene positive while the remaining eight (n=8) diarrheal samples appear as fim gene positive. Twenty four (n=24) urine samples appear as pap gene positive while the remaining nine (n=9) diarrheal samples appear as papgene positive. Fourteen (n=14) urine samples appear as hly gene positive while the remaining three (n=3) diarrheal samples appear as hly gene positive. Fifteen (n=15) urine samples appear as sfa gene positive while in diarrheal samples was not detected. The study concluded that fim gene was highly prevalent among UTIs patients, aer gene was high prevalent among diarrhea patients and Amikacin is the most effective antibiotic