DETECTION OF CO-EXISTENCE OF CARBAPENEMASE GENES USING MULTIPLEX PCR AMONG GRAM NEGATIVE BACTERIA ISOLATED FROM TERTIARY CARE HOSPITAL IN HYDERABAD
Abstract
Introduction: The emergence of carbapenem resistant organisms has been causing a grave challenge to the physicians and healthcare workers to treat the infections. Several carbapenemase genes have been reported to be involved in the development of carbapenem drug resistance. In the present study we intend to evaluate the role and presence of carbapenemase genes KPC, VIM and OXA-48 like through multiplex PCR in different gram negative bacteria.
Materials and Methods: A total of 100 bacterial isolates from the clinical samples urine -60, endotracheal secretions -18, blood culture -11, pus -8 and sputum-3. The gram negative bacteria isolates included Escherichia coli -53, Klebsiella pneumoniae-30, Pseudomonas aeruginosa- 13 and Acinetobacter Sp.-4. The organisms isolated were subjected to Kirby Bauer Antibiotic sensitivity test. The DNA extraction has been performed using bacterial DNA extraction kit (Quiagen QIAmp kit) as per the instructions of the manufacturer. The multiplex PCR has been used for the detection of KPC, VIM and OXA-48 like.
Results: The carbapenem drug resistance has been noticed in 35% of cases. The PCR has been detected VIM in 22% of isolates OXA - 48 in 20 % of isolates and KPC 13% of isolates. In 5% of cases in which the routine antibiotic sensitivity cases were showing sensitive to meropenem and imipenem has shown the presence of KPC carbapenem resistant genes.
Conclusion: Our study has shown the coexistence of multiple genes in a single bacteria pointing out that different carbapenmases enzymes are utilized by the bacteria to inactive the carbapenem drugs. The infection control practices and antibiotic stewardship will ensure the prevention of spread of carbapenem resistance.
