High Resolution Melting Curve Analysis Method for Detecting of Carbapenemases Producing Pseudomonas aeruginosa
Journal Title: Journal of Krishna Institute of Medical Sciences University - Year 2018, Vol 7, Issue 4
Abstract
Background: The, carbapenems have the broadest spectrum of activity among all β-lactam antibiotics, and also, carbapenem resistant P. aeruginosa may be susceptible to other β-lactam antibiotics. There are several methods for detecting to Carbapenems resistance of Pseudomonas aeruginosa (P. aeruginosa) strains. Aim and Objectives: In the current study, High Resolution Melt curve analysis (HRM) is evaluated to detect carbapenem resistant P. aeruginosa. Material and Methods: This experimental study was done on standard isolates of P. aeruginosa and carbapenemase producing strains in Hamadan. 16srRNA of P. aeruginosa and KPC gene of carbapenemase producing strains were applied to detect by HRM. Also, sensitivity and specificity of primers have been evaluated and carbapenemase producing P. aeruginosa were determined based on melt curve temperature range. Finally, melt curve profiles were assessed and analyzed using StepOne Software v2.3 and HRM Software v3.0.1 software. Results: According to dilutions ratios of 108 to 10-4 it was found that the designed primers have the ability to identify 10-3 CFU/ml of bacteria for P. aeruginosa and 10-1 CFU/ml for carbapenemase producing strains. The melt curve was also shown at 80.9±0/5 °C for all DNA dilutions of P. aeruginosa and 82.4±0/5 °C for all DNA dilutions of Carbapenemases strains. Conclusion: The sensitivity and specificity of HRM method indicated that it is highly reliable, rapid and cost-effective to detect carbapenemase producing P. aeruginosa.
Authors and Affiliations
Hamed Tahmasebi, Sanaz Dehbashi, Mohammad Reza Arabestani
Keywords
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