Related Experiment Video
Updated: Feb 26, 2026
Multiplex PCR Assay for Typing of Staphylococcal Cassette Chromosome Mec Types I to V in Methicillin-resistant Staphylococcus aureus
Published on: September 5, 2013
Multiplex PCR scheme for variant plasmid mediated class C β-lactamase typing
Le Van Chuong1,2, Virapong Prachayasittikul1, Chartchalerm Isarankura Na Ayudhya1
1Department of Clinical Microbiology and Applied Technology, Faculty of Medical Technology, Mahidol University, Bangkok, Thailand.
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.
Insights
A new multiplex PCR method accurately identifies and classifies diverse plasmid-mediated AmpC (pAmpC) types. This tool aids in tracking antibiotic resistance patterns and understanding pAmpC distribution globally.
Area of Science:
- Microbiology
- Molecular Biology
- Infectious Diseases
Background:
- Plasmid-mediated AmpC (pAmpC) prevalence and diversity are increasing globally.
- pAmpC contributes to beta-lactam antibiotic resistance, complicating treatment.
- Lack of standardized pAmpC identification hinders surveillance and infection control.
Purpose of the Study:
- To develop a robust method for classifying pAmpC beta-lactamases.
- To address challenges in epidemiological surveillance and infection control.
Main Methods:
- A single-tube multiplex PCR assay was designed.
- The assay classifies six distinct pAmpC groups: CIT, ECB, MOX & CMY-1 like, DHA, ACC, and FOX.
- Method validation included sensitivity and specificity testing.
Main Results:
- The developed PCR method can detect DNA at nano-scale concentrations (2.5 ng).
- It demonstrates high discriminatory power compared to phenotypic and commercial genotypic methods.
- Successfully classifies six pAmpC groups.
Conclusions:
- The multiplex PCR is a valuable tool for monitoring clinically significant resistance patterns.
- Facilitates further investigation into the occurrence and distribution of pAmpC types.
- Aids in epidemiological surveillance and infection control strategies.
