Real-Time PCR Assays compared to Culture-Based Approaches for Identification of Aerobic Bacteria in Chronic Wounds.
Clin Microbiol Infect. 2009 Dec 29;
Authors: Melendez JH, Frankel YM, An AT, Williams L, Price LB, Wang NY, Lazarus GS, Zenilman JM
ABSTRACT Chronic wounds cause substantial morbidity and disability. Infection in chronic wounds is clinically defined by routine culture methods which take several days to finalize, and may not fully describe the community of organisms or biome within these wounds. Molecular diagnostic approaches offer promise for 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 demonstrated a high degree of sensitivity and specificity against a panel of organisms commonly associated with chronic wound infection. Thirty-nine tissue samples from 29 chronic wounds were evaluated and the results compared to cultures. By culture and PCR, the most common organisms were Methicillin Resistant Staphylococcus aureus (MRSA) followed by Streptococcus agalactiae (Group B streptococcus: GBS) and Pseudomonas aeruginosa. The sensitivities of the PCR assays were 100% and 90% when quantitative and qualitative culture results were used as the gold standard, respectively. The assays identified bacterial DNA from 10 additional organisms that were not reported by quantitative or qualitative cultures. Under optimal conditions, turnaround for PCR results is as little as 4-6 hours. Real-Time PCR is a rapid and inexpensive approach, which can be easily translated 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.
PMID: 20047604 [PubMed - as supplied by publisher]