Molecular Dipstick Test for Diagnosis of Sleeping Sickness

• Published in: Journal of Clinical Microbiology 2006-08, Vol.44 (8), p.2884-2889
• Main Authors: Deborggraeve, S, Claes, F, Laurent, T, Mertens, P, Leclipteux, T, Dujardin, J.C, Herdewijn, P, Büscher, P
• Format: Article
• Language: English
• Subjects: Animals; Base Sequence; Biological and medical sciences; Blood - parasitology; DNA, Protozoan - analysis; DNA, Protozoan - genetics; DNA, Protozoan - isolation & purification; Fundamental and applied biological sciences. Psychology; Human protozoal diseases; Humans; Infectious diseases; Medical sciences; Microbiology; Molecular Diagnostic Techniques; Molecular Sequence Data; Nucleic Acid Hybridization - methods; Parasitic diseases; Parasitology; Polymerase Chain Reaction - methods; Protozoal diseases; Reference Standards; Sensitivity and Specificity; Trypanosoma brucei; Trypanosoma brucei brucei - genetics; Trypanosoma brucei brucei - isolation & purification; Trypanosomiasis; Trypanosomiasis, African - diagnosis
• ISSN: 0095-1137; 1098-660X; 1098-5530
• DOI: 10.1128/JCM.02594-05

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.