DETECTION OF CARBAPENEM AND COLISTIN RESISTANT GENES IN PRESERVED SAMPLES OF P. AERUGINOSA

Abstract

Pseudomonas aeruginosa is a rod shaped, aerobic and gram-negative bacteria that is found in humans, soil as well as water and plants. It has become worldwide threat to immunocompromised patients due to its increasing resistance profile. The aim of present research is to revive Pseudomonas aeruginosa from preserved samples having different sources, i.e. clinical, diabetic foot ulcer, raw milk, chicken embryo and broiler and cockroach samples. This study also targets antibiotic susceptibility profile of P. Aeruginosa against Carbapenem and Colistin for the detection of three resistant genes i.e blaKPC, blaIMP and MCR-1 gene. A total of 150 samples were revived, in which sixty-five samples show growth for P. aeruginosa. All these samples were enriched in Peptone water for 48 hours. All samples were cultured and analyzed on the Cetrimide agar. Among sixty-five, twenty-four (n=24) were Clinical samples, twenty (n=20) were diabetic foot ulcers, seven (n=7) were raw milk samples, ten (n=10) were chicken embryos and broiler and four (n=4) were cockroach samples. P. aeruginosa was identified based on colony morphology subsequently microscopy after the Gram-staining and biochemical tests. After identification, the bacterium was further tested for antimicrobial sensitivity according to the Kirby-Baur method. Antibiotic tests showed 100% resistance against carbapenem (imipenem, meropenem and Doripenem) and 0% Colistin resistance. None of the samples showed Colistin resistance. The DNA of the Pseudomonas aeruginosa was extracted by chemical method and ran on the gel electrophoresis. Furthermore, the DNA bands were visualized under UV Transilluminator. Extracted DNA was further amplified by using the site-specific primer of drug resistance gene blaKPC, blaIMP and mcr-1. The PCR results showed 9.23% positive isolates for KPC gene and 6.15% positive IMP gene. None of these sixty-five isolates contain mcr1 gene. In summary, our study's findings could help with the development of therapeutic strategies and the evaluation of the efficacy of currently being used antibiotic therapies. The study offers insightful details regarding profile of antibiotic resistance. The presence of KPC and IMP genes is important to track the cause of resistance in Pseudomonas aeruginosa.