MOLECULAR CHARACTERISATION OF EXO T GENE AS A VIRULENCE FACTOR IN BIOFILM PRODUCTION IN Pseudomonas aeruginosa ISOLATES AT A TERTIARY CARE CENTRE KANPUR

Abstract

Introduction: The emergence of multidrug-resistant strains of Pseudomonas aeruginosa has complicated medical therapy. Antimicrobial resistance and biofilm formation are virulence factors that contribute to the persistence of long-term infections. Gene ExoT and Exo U plays an important role in causation of infection and can be a notable virulent element expressed by P.aeruginosa clinical isolates. Aim and Objective: To Study the Molecular characterization of ExoT and ExoU Gene from Pseudomonas aeruginosa and its Antibiogram with the ability of Biofilm Formation of Patients attending a Tertiary Care Centre. Material and Methods: This was a cross sectional study conducted in the Department of Microbiology at RMCHandRC, Kanpur, Uttar Pradesh. A Total of 132 Pseudomonas aeruginosa isolated from different clinical samples including urine, sputum, ear swab, wound swab, pus were identified by standard microbiological techniques. The isolates were further tested for MBL by Imipenem EDTA combined disc test and MBL E-test (Imipenem). The DNA was isolated by Ozone DNA extraction kit as per the manufacture s guidelines and the resistant gene Exo T and Exo U was detected by the conventional PCR. Results: A total of 355 clinical samples were studied out of which 132 P. aeruginosa isolates were obtained. The prevalence rate of Pseudomonas aeruginosa was observed to be 37.1%. Sensitivities of Colistin was (95.8%), Piperacillin-tazobactam (72.9%), Amikacin (77 %), and cefepime (72.9%) were found to be the most effective Antibiotics. The resistance to ciprofloxacin was (43.7%), Levofloxacin (50%), Piperacillin(46.6%), and Gentamicin(64.5%), Imipenem (64.5%), Tobramycin(68.7%), Ceftazidime (68.7%). Of which 20(15.1%%) were MBL positive by Imipenem EDTA combined disc test, and 19(20.45%) by MBL E- test. 50.75% (n 67) of isolates showed biofilm phenotype. The molecular characterization confirms 56 (42.4%) expression exoT gene and Exo U gene was observed to be 21 (15.9%). Conclusion: In our study, MBL E-Test was found to be the best ph