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Am. J. Trop. Med. Hyg., 72(5), 2005, pp. 640-641
Copyright © 2005 by The American Society of Tropical Medicine and Hygiene

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SHORT REPORT


DETECTION OF ORIENTIA TSUTSUGAMUSHI IN CLINICAL SAMPLES BY QUANTITATIVE REAL-TIME POLYMERASE CHAIN REACTION

TASAWAN SINGHSILARAK, WATTANA LEOWATTANA, SORNCHAI LOOAREESUWAN, VAREE WONGCHOTIGUL, JU JIANG, ALLEN L. RICHARDS, AND GEORGE WATT
Department of Clinical Tropical Medicine, Faculty of Tropical Medicine, Mahidol University, Bangkok, Thailand; Department of Microbiology, Faculty of Tropical Medicine, Mahidol University, Bangkok, Thailand; Rickettsial Diseases Department, Naval Medical Research Center, Silver Spring, Maryland; Department of Clinical Tropical Medicine, Faculty of Tropical Medicine, Mahidol University, Bangkok, Thailand and Department of Medicine, Hawaii AIDS Clinical Research Program, University of Hawaii, Manoa, Honolulu, Hawaii

 

ABSTRACT

Orientia tsutsugamushi infection causes scrub typhus, a common zoonosis of rural Asia. Orientia tsutsugamushi was recently detected by a real-time quantitative polymerase chain reaction (qPCR) assay in animal specimens. We evaluated the same qPCR assay in specimens obtained from patients with serologically proven scrub typhus infections. The 47-kDa qPCR assay was more sensitive than was mouse inoculation; it was reactive in whole blood specimens from all 10 isolate-positive patients and in 7 of 17 isolate-negative individuals (P = 0.003, Fisher’s two-tailed exact test). As few as 1,076 O. tsutsugamushi copies/µL were detected in whole blood. Four of 7 sera from isolate-proven scrub typhus infections were also reactive by qPCR. The assay was unreactive in all 12 individuals without scrub typhus infection. This is the first demonstration of a sensitive and specific real-time qPCR assay for human scrub typhus infection.

Received May 26, 2004. Accepted for publication August 30, 2004.

Financial support: This work was supported by the Faculty of Tropical Medicine, Mahidol University.

Authors’ addresses: Tasawan Singhsilarak, Department of Clinical Tropical Medicine, Faculty of Tropical Medicine, Mahidol University, Bangkok, Thailand 10400, Telephone: 662-354-9100 ext. 2073, Fax: 662-354-1792, E-mail: tmtss{at}mahidol.ac.th. Wattana Leowattana, Department of Clinical Tropical Medicine, Faculty of Tropical Medicine, Mahidol University, Bangkok, Thailand 10400, Telephone: 662-354-9100 ext. 2039, Fax: 662-354-9168, E-mail: leowattana{at}yahoo.com. Sornchai Looareesuwan, Department of Clinical Tropical Medicine, Faculty of Tropical Medicine, Mahidol University, Bangkok, Thailand 10400, Telephone: 662-354-9159, Fax: 662-354-9158, E-mail: tmslr{at}mshidol.ac.th. Varee Wongchotigul, Department of Microbiology, Faculty of Tropical Medicine, Mahidol University, Bangkok, Thailand 10400, Telephone: 662-354-9100 ext. 2075, E-mail: tmvwc{at}mahidol.ac.th. Ju Jiang, Rickettsial Diseases Department, Naval Medical Research Center, 503 Robert Grant Avenue, Silver Spring, MD 20910-7500, Telephone: 301-319-7249, Fax: 301-319-7460, E-mail: JiangJ{at}nmrc.navy.mail. Allen L. Richards, Rickettsial Diseases Department, Naval Medical Research Center, 503 Robert Grant Avenue, Silver Spring, MD 20910-7500, Telephone: 301-319-7668, Fax: 301-319-7460, E-mail: RichardsA{at}nmrc.navy.mil. George Watt, Department of Clinical Tropical Medicine, Faculty of Tropical Medicine, Mahidol University, Bangkok, Thailand 10400, Telephone: 662-354-1754, Fax: 662-354-1792, E-mail: gwattth{at}yahoo.com.

Reprint requests: George Watt, Department of Clinical Tropical Medicine, Faculty of Tropical Medicine, Mahidol University, Bangkok, Thailand 10400.




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