Real-time PCR assays compared to culture-based approaches for identification of aerobic bacteria in chronic wounds

J. H. Melendez, Y. M. Frankel, A. T. An, L. Williams, L. B. Price, Nae Yuh Wang, Gerald Sylvan Lazarus, Jonathan Mark Zenilman

Research output: Contribution to journalArticle

Abstract

Chronic wounds cause substantial morbidity and disability. Infection in chronic wounds is clinically defined by routine culture methods that can take several days to obtain a final result, and may not fully describe the community of organisms or biome within these wounds. Molecular diagnostic approaches offer promise for a more rapid and complete assessment. We report the development of a suite of real-time PCR assays for rapid identification of bacteria directly from tissue samples. The panel of assays targets 14 common, clinically relevant, aerobic pathogens and demonstrates a high degree of sensitivity and specificity using a panel of organisms commonly associated with chronic wound infection. Thirty-nine tissue samples from 29 chronic wounds were evaluated and the results compared with those obtained by culture. As revealed by culture and PCR, the most common organisms were methicillin-resistant Staphylococcus aureus (MRSA) followed by Streptococcus agalactiae (Group B streptococcus) and Pseudomonas aeruginosa. The sensitivities of the PCR assays were 100% and 90% when quantitative and qualitative culture results were used as the reference standard, respectively. The assays allowed the identification of bacterial DNA from ten additional organisms that were not revealed by quantitative or qualitative cultures. Under optimal conditions, the turnaround time for PCR results is as short as 4-6 h. Real-time PCR is a rapid and inexpensive approach that can be easily introduced into clinical practice for detection of organisms directly from tissue samples. Characterization of the anaerobic microflora by real-time PCR of chronic wounds is warranted.

Original languageEnglish (US)
Pages (from-to)1762-1769
Number of pages8
JournalClinical Microbiology and Infection
Volume16
Issue number12
DOIs
StatePublished - Dec 2010

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Aerobic Bacteria
Real-Time Polymerase Chain Reaction
Wounds and Injuries
Streptococcus agalactiae
Polymerase Chain Reaction
Bacterial DNA
Molecular Pathology
Wound Infection
Methicillin-Resistant Staphylococcus aureus
Streptococcus
Pseudomonas aeruginosa
Ecosystem
Morbidity
Bacteria
Sensitivity and Specificity
Infection

Keywords

  • Bacteria
  • Chronic wounds
  • Cultures
  • Infection
  • Real-time PCR

ASJC Scopus subject areas

  • Microbiology (medical)
  • Infectious Diseases

Cite this

Real-time PCR assays compared to culture-based approaches for identification of aerobic bacteria in chronic wounds. / Melendez, J. H.; Frankel, Y. M.; An, A. T.; Williams, L.; Price, L. B.; Wang, Nae Yuh; Lazarus, Gerald Sylvan; Zenilman, Jonathan Mark.

In: Clinical Microbiology and Infection, Vol. 16, No. 12, 12.2010, p. 1762-1769.

Research output: Contribution to journalArticle

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