Direct detection of pathogens in osteoarticular infections by polymerase chain reaction amplification and microarray hybridization.

Abstract

BACKGROUND:

Molecular biological techniques such as the polymerase chain reaction (PCR) and DNA microarray are used for the detection/identification of microorganisms; however, few reports have discussed the clinical utility of microarray analysis for identification of causative organisms of osteoarticular infections. It is important to examine the utility of PCR amplification followed by analysis of DNA microarray carrying specific oligonucleotides.

METHODS:

This study included 101 biological samples obtained from 96 patients who underwent conservative and/or surgical treatment for osteoarticular infections. In this double-blind comparative study, routine conventional testing and the research groups were unaware of each other's interpretation until identical specimens were identified by culture and microarray analysis.

RESULTS:

Results of PCR microarray analysis were positive for 25 samples and negative for the remaining 76 samples within 24 h, and the results of the cultures (available after a mean of 3.54 days) were positive in 26 samples and negative for the remaining 75 samples. The sensitivity of microarray analysis was 84.6% (22/26) and specificity was 88.0% (22/25). Discrepant results were identified in seven samples, including a negative culture and a positive microarray in three cases and a positive culture and a negative microarray in four other cases.

CONCLUSIONS:

The PCR microarray analysis is complementary to routine cultures in identifying causative microorganisms and should be used in patients with highly suspected infections and negative bacterial culture and in patients who require prompt diagnosis and early initiation of antibiotic therapy.

• Authors: Kenzo Uchida, Takafumi Yayama, Yasuo Kokubo, Tsuyoshi Miyazaki, Hideaki Nakajima, Kohei Negoro, Kenichi Takeno, Elisa S Mwaka, Norbert T Orwotho, Mitsunobu Shimadzu, Shigeru Kobayashi, Hisatoshi Baba
• Journal: Journal of orthopaedic science : official journal of the Japanese Orthopaedic Association (J Orthop Sci) Vol. 14 Issue 5 Pg. 471-83 (Sep 2009) ISSN: 1436-2023 [Electronic] Japan
• PMID: 19802657 (Publication Type: Comparative Study, Journal Article)

Chemical References

• DNA Probes

Topics

• Bone Diseases, Infectious (microbiology)
• DNA Probes
• Double-Blind Method
• Humans
• Oligonucleotide Array Sequence Analysis
• Polymerase Chain Reaction (methods)
• Sensitivity and Specificity