The use of high-resolution melting analysis for rapid spa typing on methicillin-resistant Staphylococcus aureus clinical isolates

J Microbiol Methods. 2013 Feb 15;92(2):99-102. doi: 10.1016/j.mimet.2012.11.006. Epub 2012 Nov 13.

Abstract

Methicillin-resistant Staphylococcus aureus (MRSA) has been endemic in Hong Kong for three decades. This study evaluated the practical use of high-resolution melting (HRM) real-time PCR analysis on MRSA staphylococcal Protein A (spa) typing on local MRSA isolates. Among 55 clinical MRSA isolates collected in 2011, 12 different spa types were observed by the conventional PCR-sequencing method including the locally predominant spa type t1081 and two locally predominant community acquired MRSA spa types t019 and t437. By using the HRM method, it could differentiate all 12 spa genotypes by distinct melting curves and HRM difference plot analysis. These two methods demonstrated 100% concordance whereas the HRM method required only 3h of turnaround time and one-fifth of reagent cost compared to the conventional method. Our study confirmed that the cost effective and rapid HRM typing approach is practically useful for MRSA community transmission monitoring and nosocomial outbreak control in Hong Kong.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Adolescent
  • Adult
  • Aged
  • Aged, 80 and over
  • Child
  • Community-Acquired Infections / epidemiology
  • Community-Acquired Infections / microbiology
  • Cross Infection / epidemiology
  • Cross Infection / microbiology
  • Female
  • Hong Kong
  • Humans
  • Male
  • Methicillin-Resistant Staphylococcus aureus / classification*
  • Methicillin-Resistant Staphylococcus aureus / genetics*
  • Middle Aged
  • Molecular Epidemiology / methods
  • Molecular Typing / economics
  • Molecular Typing / methods*
  • Real-Time Polymerase Chain Reaction / economics
  • Real-Time Polymerase Chain Reaction / methods*
  • Staphylococcal Infections / epidemiology
  • Staphylococcal Infections / microbiology
  • Staphylococcal Protein A / genetics
  • Transition Temperature
  • Young Adult

Substances

  • Staphylococcal Protein A