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Molecular dipstick test for diagnosis of sleeping sickness.

Abstract
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 microl 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.
AuthorsS Deborggraeve, F Claes, T Laurent, P Mertens, T Leclipteux, J C Dujardin, P Herdewijn, P Büscher
JournalJournal of clinical microbiology (J Clin Microbiol) Vol. 44 Issue 8 Pg. 2884-9 (Aug 2006) ISSN: 0095-1137 [Print] United States
PMID16891507 (Publication Type: Evaluation Studies, Journal Article, Research Support, Non-U.S. Gov't)
Chemical References
  • DNA, Protozoan
Topics
  • Animals
  • Base Sequence
  • Blood (parasitology)
  • DNA, Protozoan (analysis, genetics, isolation & purification)
  • Humans
  • Molecular Diagnostic Techniques
  • Molecular Sequence Data
  • Nucleic Acid Hybridization (methods)
  • Polymerase Chain Reaction (methods)
  • Reference Standards
  • Sensitivity and Specificity
  • Trypanosoma brucei brucei (genetics, isolation & purification)
  • Trypanosomiasis, African (diagnosis)

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