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Updated: Dec 27, 2025

Multiplex PCR Assay for Typing of Staphylococcal Cassette Chromosome Mec Types I to V in Methicillin-resistant Staphylococcus aureus

Multiplex PCR Assay for Typing of Staphylococcal Cassette Chromosome Mec Types I to V in Methicillin-resistant Staphylococcus aureus

Published on: September 5, 2013

Updated emm-typing protocol for Streptococcus pyogenes.

H R Frost¹, M R Davies², S Velusamy³

¹Molecular Bacteriology Laboratory, Université Libre de Bruxelles, Brussels, Belgium; Tropical Diseases Research Group, Murdoch Children's Research Institute, Melbourne, Australia.

Clinical Microbiology and Infection : the Official Publication of the European Society of Clinical Microbiology and Infectious Diseases

|March 3, 2020

Summary

This summary is machine-generated.

A new primer, CDC3, improves emm gene typing for Streptococcus pyogenes. This reduces misclassification of strains by ensuring PCR reactions amplify only the target emm gene sequence.

Keywords:

Group A Streptococcus M protein M-like protein PCR Streptococcus pyogene Vaccine emm-typing protocol

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Published on: July 22, 2011

Area of Science:

Microbiology
Molecular Biology
Genetics

Background:

Streptococcus pyogenes emm gene typing via PCR can lead to misclassification due to multiple band amplification.
Non-emm gene sequences have been mistakenly identified as emm types.

Purpose of the Study:

To enhance the specificity of emm typing PCR for Streptococcus pyogenes.
To introduce and validate a new primer, CDC3, for improved emm gene detection.

Main Methods:

In silico validation of the CDC3 primer against 1688 GAS genomes.
In vitro testing of CDC3 with 32 Streptococcus pyogenes isolates using PCR.
Comparison of PCR results using CDC1+CDC2 versus CDC1+CDC3 primer pairs, followed by gel electrophoresis and sequencing.

Main Results:

The CDC3 primer demonstrated high specificity, annealing only to emm genes in silico, unlike the CDC2 primer which annealed to non-emm genes.
PCR using the CDC1+CDC3 primer pair produced a single, correct-sized band in all tested isolates.
The CDC1+CDC2 primer pair produced multiple bands in 40% of isolates, indicating potential misclassification.

Conclusions:

The CDC3 primer offers superior specificity for emm gene detection in Streptococcus pyogenes compared to the CDC2 primer.
This new primer provides a straightforward method to prevent the mistyping of Streptococcus pyogenes strains.