Analysis of Genetic Diversity of Escherichia Coli Isolates Using RAPD PCR Technique

Abstract

RAPD-Polymerase chain reaction technique was used to analyze genetic variety for nine isolates of Escherichia coli, causing Urinary Tract Infection in pregnant women. Fifty-three urine samples were collected from inpatients and identified fourteen isolates of E. coli using biochemical analysis. E. coli isolates were subjected to phylogenetic analysis using fifteen random ten nucleotide primers. DNA amplification with each of the fifteen primers resulted in generation of different DNA fingerprinting profiles with a varied number of bands. The results of the reactions of RAPD technique for the nine selected isolates revealed differences of the number of amplified bands and in the molecular size of the produced bands. The total number of polymorphic bands were 122. The highest number of bands ( 13 bands ) were produced by primer A14, while the primer O20 gave the lowest number of polymorphic bands(5 bands). Dendrogram based generation of clustering of E. coli isolates showed three major clusters, members in one of which showed the most close genetic profile and biochemical properties as well. Application of Random Amplification of Polymorphic DNA as a new alternative approach in molecular characterization of E. coli infections is remarkable to trace genetic relatedness in a short duration, hence dramatically increasing its clinical relevance over existing biochemical or biogramatic methods in studying epidemiological patterns of pathogen sources and distribution manners.