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Prospective Clinical Evaluation of the Accuracy of 16S rRNA Real-Time PCR Assay for the Diagnosis of Melioidosis

The accuracy of a Burkholderia pseudomallei 16s rRNA real-time PCR assay was evaluated against culture for the diagnosis of melioidosis in Thailand. A total of 846 samples were obtained from 383 patients with suspected melioidosis. One or more specimens were PCR positive for 47 of 77 patients with culture-proven melioidosis (sensitivity 61.0%, 95% CI: 49.2–72.0%). PCR was negative for all 306 patients who were culture negative for B. pseudomallei (specificity 100%, 95% CI: 98.8–100%). Diagnostic sensitivity of PCR was 22.7% for patients who were culture positive for blood only, compared with 79.4% for patients who were culture positive for samples other than blood. The median (interquartile range) B. pseudomallei colony count in blood for 44 of 77 patients with positive blood cultures was 2.4 CFU/ml (0.2–13.5 CFU/ml); this may explain the low sensitivity of PCR for this specimen. The PCR assay described here is not sufficiently sensitive to replace culture in our clinical setting.

Received July 10, 2007. Accepted for publication August 15, 2007.

Acknowledgments: We are grateful for the support of the medical, nursing, and laboratory staff at Sappasithiprasong Hospital and staff at the Wellcome Trust-Mahidol University-Oxford University Tropical Medicine Research Program, including Gumphol Wongsuvan, Nongluk Getchalarat, Jintana Suwannapreuk, Sukallaya Paengmee, and Nittaya Teerawattanasook.

Financial support: This study was funded by the Wellcome Trust. SJP was supported by a Wellcome Trust Career Development Award in Clinical Tropical Medicine.

^* Address correspondence to Sharon Peacock, Mahidol-Oxford Tropical Medicine Research Unit, Faculty of Tropical Medicine, Mahidol University, 420/6 Rajvithi Road, Bangkok, 10400, Thailand. E-mail: sharon{at}tropmedres.ac

Authors’ addresses: Narisara Chantratita, Vanaporn Wuthiekanun, Direk Limmathurotsakul, Aunchalee Thanvisai, Nicholas P. J. Day, and Sharon J. Peacock, Mahidol-Oxford Tropical Medicine Research Unit, Faculty of Tropical Medicine, Mahidol University, 420/6 Rajvithi Road, Bangkok, 10400, Thailand, Telephone: +66 2 354 9172, Fax: +66 2 354 9169, E-mail: narisara{at}tropmedres.ac. Wasun Chantratita, Department of Pathology, Faculty of Medicine (Ramathibodi Hospital), Mahidol University, Rama VI Road, Payathai, Bangkok 10400, Thailand.

Reprint requests: Sharon Peacock, Mahidol-Oxford Tropical Medicine Research Unit, Faculty of Tropical Medicine, Mahidol University, 420/6 Rajvithi Road, Bangkok, 10400, Thailand. Tel: +66 2 354 9172, Fax: +66 2 354 9169, E-mail: sharon{at}tropmedres.ac.