1.Optimization of PCR for rapid detection of CTX-M gene in ESBL producing Klebsiella pneumoniae clinical isolates from Punjab, Pakistan
Rasheeda Bashir ; Neelam Zaib ; Imran Altaf ; Faiza Saleem ; Qamar Sultana ; Shagufta Naz
Malaysian Journal of Microbiology 2016;12(5):365-369
Aims: The study was carried out firstly, to evaluate the prevalence of extended spectrum beta lactamase (ESBL),
multidrug resistant Klebsiella pneumoniae isolates from Punjab, Pakistan and secondly, to characterize the genotypes of
their beta lactamase producing enzymes and optimization of PCR based method for rapid and authentic detection of
antibiotic resistant gene.
Methodology and results: Two hundred of K. pneumonia strains were isolated from different clinical samples. Blood
and MacConkey Agar were used to isolate and identify bacterial microorganisms while Muller Hinton Agar was used to
evaluate the antimicrobial susceptibility against different antibiotics as per CLSI 2012 guidelines. ESBL producing
bacteria were screened by double disk synergy /combination disk test. PCR was optimized and performed for resistant
gene (CTX-M). The results showed that most of the isolates were resistant to multiple antibiotic including cephalosporin,
aztreonam, sulphamethoxazole/trimethoprim, ciprofloxacin, doxycyclin and were sensitive to imipenam and amikacin.
Frequency of CTX-M gene in ESBL producing K. pneumoniae was 94%.
Conclusion, significance and impact of study: Based on the finding of this study it is suggested that prevalence of
CTX-M gene (95%) is very high among ESBL producing isolates. Therefore, PCR based method may help clinicians for
rapid detection and treatment of patients by choosing right medication against the resistant bacteria as early as possible.