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Journal of Clinical Microbiology, August 2006, p. 2884-2889, Vol. 44, No. 8
0095-1137/06/$08.00+0     doi:10.1128/JCM.02594-05
Copyright © 2006, American Society for Microbiology. All Rights Reserved.

Molecular Dipstick Test for Diagnosis of Sleeping Sickness

S. Deborggraeve,1,3* F. Claes,1 T. Laurent,1 P. Mertens,2 T. Leclipteux,2 J. C. Dujardin,1 P. Herdewijn,3 and P. Büscher1

Department of Parasitology, Institute of Tropical Medicine, Antwerp, Belgium,1 Coris BioConcept, Gembloux, Belgium,2 Laboratory of Medicinal Chemistry, Rega Institute for Medicinal Research, Leuven, Belgium3

Received 14 December 2005/ Returned for modification 27 January 2006/ Accepted 2 June 2006

Human African trypanosomiasis (HAT) or sleeping sickness is a neglected disease that affects poor rural populations across sub-Saharan Africa. Confirmation of diagnosis is based on detection of parasites in either blood or lymph by microscopy. Here we present the development and the first-phase evaluation of a simple and rapid test (HAT-PCR-OC [human African trypanosomiasis-PCR-oligochromatography]) for detection of amplified Trypanosoma brucei DNA. PCR products are visualized on a dipstick through hybridization with a gold-conjugated probe (oligochromatography). Visualization is straightforward and takes only 5 min. Controls both for the PCR and for DNA migration are incorporated into the assay. The lower detection limit of the test is 5 fg of pure T. brucei DNA. One parasite in 180 µl of blood is still detectable. Sensitivity and specificity for T. brucei were calculated at 100% when tested on blood samples from 26 confirmed sleeping sickness patients, 18 negative controls (nonendemic region), and 50 negative control blood samples from an endemic region. HAT-PCR-OC is a promising new tool for diagnosis of sleeping sickness in laboratory settings, and the diagnostic format described here may have wider application for other infectious diseases.

* Corresponding author. Mailing address: Department of Parasitology, Institute of Tropical Medicine, Nationalestraat 155, B-2000 Antwerpen, Belgium. Phone: 32 3 24 76 488. Fax: 32 3 24 76 359. E-mail: sdeborggraeve{at}itg.be.

Journal of Clinical Microbiology, August 2006, p. 2884-2889, Vol. 44, No. 8
0095-1137/06/$08.00+0     doi:10.1128/JCM.02594-05
Copyright © 2006, American Society for Microbiology. All Rights Reserved.



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