A PCR method for the identification of methicillin-resistant Staphylococcus aureus (MRSA) from screening swabs

Pathology. 2004 Jun;36(3):265-8. doi: 10.1080/00313020410001692530.

Abstract

Aim: The aim of this study was to assess the discriminatory power and potential turn around time (TAT) of a PCR-based method for the detection of methicillin-resistant Staphylococcus aureus (MRSA) from screening swabs.

Methods: Screening swabs were examined using the current laboratory protocol of direct culture on mannitol salt agar supplemented with oxacillin (MSAO-direct). The PCR method involved pre-incubation in broth for 4 hours followed by a multiplex PCR with primers directed to mecA and nuc genes of MRSA. The reference standard was determined by pre-incubation in broth for 4 hours followed by culture on MSAO (MSAO-broth).

Results: A total of 256 swabs was analysed. The rates of detection of MRSA using MSAO-direct, MSAO-broth and PCR were 10.2, 13.3 and 10.2%, respectively. For PCR, the sensitivity, specificity, positive predictive value and negative predictive values were 66.7% (95%CI 51.9-83.3%), 98.6% (95%CI 97.1-100%), 84.6% (95%CI 76.2-100%) and 95.2% (95%CI 92.4-98.0%), respectively, and these results were almost identical to those obtained from MSAO-direct. The agreement between MSAO-direct and PCR was 61.5% (95%CI 42.8-80.2%) for positive results, 95.6% (95%CI 93.0-98.2%) for negative results and overall was 92.2% (95%CI 88.9-95.5%).

Conclusions: (1) The discriminatory power of PCR and MSAO-direct is similar but the level of agreement, especially for true positive results, is low. (2) The potential TAT for the PCR method provides a marked advantage over conventional methods. (3) Further modifications to the PCR method such as increased broth incubation time, use of selective broth and adaptation to real-time PCR may lead to improvement in sensitivity and TAT.

Publication types

  • Comparative Study
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Bacterial Proteins / genetics
  • Bacterial Typing Techniques*
  • Culture Media
  • DNA Primers
  • Endonucleases / genetics
  • Humans
  • Mass Screening
  • Methicillin Resistance / genetics*
  • Micrococcal Nuclease / genetics
  • Penicillin-Binding Proteins
  • Polymerase Chain Reaction / methods*
  • Sensitivity and Specificity
  • Staphylococcal Infections / microbiology*
  • Staphylococcus aureus / isolation & purification*

Substances

  • Bacterial Proteins
  • Culture Media
  • DNA Primers
  • Penicillin-Binding Proteins
  • mecA protein, Staphylococcus aureus
  • nuc protein, staphylococcus
  • Endonucleases
  • Micrococcal Nuclease