Identification of group A streptococcal emm types commonly associated with invasive infections and antimicrobial resistance by the use of multiplex PCR and high-resolution melting analysis

Eur J Clin Microbiol Infect Dis. 2012 Oct;31(10):2817-26. doi: 10.1007/s10096-012-1635-5. Epub 2012 May 20.

Abstract

M/emm typing, based either on serotyping of the M protein or on sequencing of the emm gene, is a major tool for epidemiological studies of group A streptococci (GAS). In order to simplify M/emm typing, we designed two multiplex polymerase chain reaction (PCR) formats capable of identifying the most frequent GAS M/emm types involved in invasive infections and antimicrobial resistance. A heptaplex PCR procedure was first developed in a conventional format coupled with gel electrophoresis to identify emm types 1, 3, 4, 6, 12, 28, and 89, based on the size of the amplification products. The other method, designed to identify the same seven emm types, together with emm11, was based on a real-time PCR format coupled with high-resolution melting (HRM) analysis, allowing the rapid typing of large strain collections.

MeSH terms

  • Adolescent
  • Anti-Infective Agents / pharmacology
  • Antigens, Bacterial / analysis*
  • Bacterial Outer Membrane Proteins / analysis*
  • Bacterial Typing Techniques / methods*
  • Carrier Proteins / analysis*
  • Child
  • Child, Preschool
  • DNA, Bacterial / analysis
  • Drug Resistance, Bacterial*
  • Electrophoresis, Agar Gel / methods
  • Erythromycin / pharmacology
  • Genes, Bacterial
  • Humans
  • Infant
  • Infant, Newborn
  • Multiplex Polymerase Chain Reaction / methods*
  • Nucleic Acid Denaturation
  • Reproducibility of Results
  • Sensitivity and Specificity
  • Streptococcal Infections / blood
  • Streptococcal Infections / epidemiology
  • Streptococcal Infections / microbiology*
  • Streptococcus / classification
  • Streptococcus / drug effects
  • Streptococcus / genetics
  • Streptococcus / isolation & purification*
  • Time Factors

Substances

  • Anti-Infective Agents
  • Antigens, Bacterial
  • Bacterial Outer Membrane Proteins
  • Carrier Proteins
  • DNA, Bacterial
  • streptococcal M protein
  • Erythromycin