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Updated: Mar 8, 2026
Multiplex PCR and Reverse Line Blot Hybridization Assay mPCR/RLB
Published on: August 6, 2011
Rapid discrimination of blaIMP-1, blaIMP-6, and blaIMP-34 using a multiplex PCR
Shizuo Kayama1, Hiroki Ohge2, Motoyuki Sugai1
1Project Research Center for Nosocomial Infectious Diseases, Hiroshima University, Japan; Department of Bacteriology, Hiroshima University Graduate School of Biomedical and Health Sciences, Japan.
Abstract:
Stealth-type carbapenem-resistant Enterobacteriaceae that are resistant to almost all β-lactams except imipenem is emerging in Japan. This resistance is mediated by specific variants of the metallo-β-lactamases (blaIMP-6 or blaIMP-34) that differs by one amino acid from the common variant blaIMP-1. We developed an amplification refractory mutation system (ARMS) PCR assay enabling rapid, sequence independent, identification of these variants.
Insights
Emerging stealth-type carbapenem-resistant Enterobacteriaceae in Japan are resistant to most beta-lactams. A new ARMS PCR assay rapidly identifies specific metallo-beta-lactamase variants responsible for this resistance.
Area of Science:
- Microbiology
- Molecular Biology
- Infectious Diseases
Background:
- Carbapenem-resistant Enterobacteriaceae (CRE) pose a significant global health threat.
- Stealth-type CRE, resistant to most beta-lactams except imipenem, are emerging in Japan.
- This resistance is linked to specific metallo-beta-lactamase (MBL) variants, blaIMP-6 and blaIMP-34.
Purpose of the Study:
- To develop a rapid and accurate method for identifying emerging stealth-type CRE.
- To differentiate between common and novel MBL variants mediating carbapenem resistance.
Main Methods:
- Development of an Amplification Refractory Mutation System (ARMS) PCR assay.
- The assay is designed for sequence-independent identification of blaIMP-6 and blaIMP-34 variants.
- Validation of the assay for rapid detection.
Main Results:
- Successful development of an ARMS PCR assay for detecting specific MBL variants.
- The assay enables rapid, sequence-independent identification of blaIMP-6 and blaIMP-34.
- This method facilitates the tracking of emerging stealth-type CRE.
Conclusions:
- The developed ARMS PCR assay is a valuable tool for the surveillance of emerging carbapenem resistance.
- Rapid identification of these MBL variants can aid in infection control strategies.
- This technology supports timely clinical and public health responses to antimicrobial resistance.
