Systematic PCR detection in culture-negative osteoarticular infections.

Abstract

BACKGROUND:

Identification of microorganisms is crucial for the successful treatment of osteoarticular infections. Molecular methods are more sensitive than culture-dependent methods but may suffer from lack of specificity.

METHODS:

We studied a large series of 3840 bone and joint culture-negative samples collected from 2308 patients hospitalized in Marseille University Hospitals from November 2007 to October 2009. The samples were systematically cultured for 15 days, and conventional broad-range polymerase chain reaction (PCR) (16S rDNA and 18S rDNA) as well as real-time PCR assays targeting human Bglobin, Staphylococcus aureus, and Kingella kingae were realized on one culture-negative specimen.

RESULTS:

Specimens from 741 patients (32.1%) tested positive by culture, including 38 in which bacteria grew only after 6 days of incubation. PCR was positive in 141 (9%) culture-negative specimens. Microorganisms identified by PCR were classified into 2 groups: fastidious bacteria (n = 35), mostly anaerobes in adult patients, and K. kingae in children; and nonfastidious bacteria (n = 106), mostly S. aureus (32.7%). A discrepancy between a positive PCR result for S. aureus and a negative culture were explained by previous antibiotherapy in 31.4% of cases. Our study highlights the usefulness of systematic 16S rDNA gene PCR for the diagnosis of bone and joint infections in culture-negative patients, thus enabling the administration of specific antibiotic treatments.

CONCLUSIONS:

We recommend the use of conventional broad-range PCR for culture-negative bone and joint specimens, as well as S. aureus-specific PCR for adults and K. kingae-specific PCR for children. 18S rDNA PCR should be reserved only for specific cases.

• Authors: Pierre-Yves Levy, Pierre-Edouard Fournier, Florence Fenollar, Didier Raoult
• Journal: The American journal of medicine (Am J Med) Vol. 126 Issue 12 Pg. 1143.e25-33 (Dec 2013) ISSN: 1555-7162 [Electronic] United States
• PMID: 24135511 (Publication Type: Journal Article, Research Support, Non-U.S. Gov't)
• Copyright: Copyright © 2013 Elsevier Inc. All rights reserved.

Chemical References

• DNA, Bacterial
• RNA, Bacterial
• RNA, Ribosomal, 16S

Topics

• Adult
• Bacteria (classification, isolation & purification)
• Bacterial Infections (diagnosis)
• DNA, Bacterial (genetics, isolation & purification)
• Humans
• Joint Diseases (diagnosis, microbiology)
• Polymerase Chain Reaction (methods)
• RNA, Bacterial (genetics, isolation & purification)
• RNA, Ribosomal, 16S (genetics)
• Retrospective Studies