Antibiotic Susceptibility Test of Klebsiella pneumoniae and K. oxytoca Isolated from Different Clinical Samples and Perform Random Amplified Polymorphic DNA among K. pneumoniae

Aims: To study the prevalence of MDR and ESBL producing Klebsiella pneumoniae and Klebsiella oxytocaand perform RAPD (Random amplified polymorphic DNA) by optimizing PCR among isolated ESBL producing Klebsiella pneumoniae.

Place and Duration of Study: This study was done to assess the prevalence of MDR and ESBL producing Klebsiella pneumoniae and K. oxytoca in urine, pus and sputum from March 2013 to April 2014 at KIST Medical College and PCR was performed at Nepal Academy of Science and Technology upto March, 2015.

Methodology: K. pneumoniae and K. oxytoca were isolated from urine, pus and sputum samples from KIST Medical College, Lalitpur, Nepal. Antibiotic susceptibility test was performed by using disk diffusion method. MDR isolates which were suspected as ESBL producers were confirmed by using double disk diffusion test and combined disk diffusion test for same isolates. Chromosomal DNA was isolated from ESBL producing K. pneumoniae. PCR was optimized by varying different reagents and visualized using gel electrophorosis. Under optimized condition chromosomal DNA was amplified, gel electrophoresis was performed and polymorphism was detected.

Results: The drug resistance pattern of K. oxytoca was high as compared to K. pneumoniae. In urine, pus and sputum samples the growth of both organisms was very much low i.e. 2.5% among total samples. 90% of K. pneumoniae isolated from urine were ESBL producers whereas only 42.85% of K. pneumoniae from pus and sputum were ESBL producers. K. oxytoca isolated from all samples were MDR as well as ESBL producers. Chromosomal DNA analysis of different antibiotic resistance pattern among K. pneumoniaeshowed higher the drug resistance lowers the polymorphism.

Conclusions: This study showed that higher the drug resistance lesser the polymorphism and greater the adaptability towards used antibiotics by production of antibiotic hydrolyzing enzymes. So, further study is required for the determination of polymorphism and drug resistance pattern correlation with organisms' adaptability to validate this concept.