Comparative studies on different molecular methods for epidemiological typing of multi-drug-resistance Klebsiella pneumoniae isolated from Mansoura hospitals

Abstract

The present study aims to evaluate two molecular methods for epidemiological typing of multi drug resistant Klebsiella pneumoniae isolated from Mansoura Hospitals. In this study, a total of 300 clinical isolates were collected from different patients distributed among Mansoura Hospitals, Dakahlia governorate, Egypt. Ninety six isolates were identified as K. pneumoniae using standard biological methods. Most isolates were obtained from urinary tract infection (70%). The susceptibility to eleven antimicrobials was determined and analyzed among isolates. The antimicrobial susceptibility test showed that Forty five isolates (47%) were multi drug resistance (MDR). Plasmids isolation from all MDR isolates revealed that 80% of these isolates harbored plasmids. Thirteen plasmid patterns were observed upon single digestion with EcoR1. However, the absence of plasmids from 20% of isolates decreases the type ability power of this technique. Hence, Random Amplified Polymorphic DNA (RAPD) analysis was applied as another molecular typing method using two individual primers, AP3 and OPA13. OPA13 showed eight distinct patterns (RA1-RA8) representing 37 isolates. Primer AP3 showed more discriminatory patterns (R1-R18) and can be used individually for better interpretation in typing a large number of isolates. In conclusion, RAPD typing is efficient and cost-effective while maintaining reproducible results for analyzing large numbers of clinical isolates and useful to understand the distribution of K. pneumoniae during investigation of outbreaks.