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

© 2017 Wiley Periodicals, Inc. Background: An increasing of prevalence and diversification of plasmid-mediated AmpC (pAmpC) has been emerged worldwide. The incidence of pAmpC resulted in increasing β-lactamase production and conferred resistance to almost all β-lactam antibiotics excluding carbapene...

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Bibliographic Details
Main Authors: Le Van Chuong, Virapong Prachayasittikul, Chartchalerm Isarankura Na Ayudhya, Ratana Lawung
Other Authors: Mahidol University
Format: Article
Published: 2019
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Online Access:https://repository.li.mahidol.ac.th/handle/123456789/45221
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Summary:© 2017 Wiley Periodicals, Inc. Background: An increasing of prevalence and diversification of plasmid-mediated AmpC (pAmpC) has been emerged worldwide. The incidence of pAmpC resulted in increasing β-lactamase production and conferred resistance to almost all β-lactam antibiotics excluding carbapenems. The lack of standard method for pAmpC identification and classification exert a challenge in epidemiological surveillance and infection control practices. Methods: A 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. Results: Developed method can detect crude extracted DNA template at nano-scale (2.5 ηg) and has high discriminatory power as compared to phenotypic and commercial genotypic method. Conclusion: The 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.