Multiplex PCR scheme for variant plasmid mediated class C β-lactamase typing.

BackgroundAn increasing of prevalence and diversification of plasmid-mediated AmpC (pAmpC) has been emerged worldwide. The incidence of pAmpC resulted in increasing -lactamase production and conferred resistance to almost all -lactam antibiotics excluding carbapenems. The lack of standard method for pAmpC identification and classification exert a challenge in epidemiological surveillance and infection control practices. MethodsA robust, single tube multiplex PCR has been developed to classify six different pAmpC groups including CIT (CMY-2 like, LAT and CFE), ECB (ACT, MIR), MOX & CMY-1 like, DHA, ACC, and FOX. The developed method was optimized and validated by testing of sensitivity and specificity. ResultsDeveloped method can detect crude extracted DNA template at nano-scale (2.5 g) and has high discriminatory power as compared to phenotypic and commercial genotypic method. ConclusionThe developed method can be utilized for tracking the changes of clinically important resistance patterns and further investigation of occurrence and distribution of plasmid-mediated AmpC types.