Phenotypic Identification and Detection of BlaCTX and TetM in Klebsiella Species from Urine Samples of Patients in Ile-Ife, Nigeria

Abstract

Klebsiella species from urine samples patients in Ile-Ife were investigated. Isolation, characterization and identification of the isolates were done using standard microbiological techniques. Antibiotic sensitivity of the Klebsiella isolates to twelve different antibiotics was determined by Kirby-Bauer’s disc diffusion method on Mueller-Hinton agar plates. Plasmid DNA in representative multiple antibiotic resistant isolates was detected by alkaline lysis (TENS buffer). Resistance (bla CTX, tet M) genes were detected by Polymerase Chain Reaction (PCR). Statistical analysis of data obtained was done using SPSS-17.0. A total of thirty-two Klebsiella sp. (7.02%) were isolated from 456 urine samples of patients; 50 urine samples collected from apparently healthy individuals served as control. Antibiotic resistance was high and also varied among the isolates with a range of 37.5% to 100%. Most of the isolates had multiple antibiotic resistance (MAR) to at least three different classes of antibiotics. There was diversity in the MAR patterns with 21 different antibiotypes which ranged from two to ten different antibiotics. Plasmid DNA of molecular weight of 900 bp to 9416 bp was detected in some representative multiple antibiotic resistant isolates. Three isolates harboured bla CTX (543 bp) gene which codes for resistance to beta-lactams. However, tet M (974 bp) gene which codes for resistance to tetracycline was not detected in any of the isolates. The study concluded that there were multiple antibiotic resistant Klebsiella species from urine samples of patients which harboured plasmid DNA and resistance (bla CTX) gene which is of great health and economic consequences.