Simplified molecular detection of Leishmania parasites in various clinical samples from patients with leishmaniasis.
BACKGROUND
Molecular methods to detect Leishmania parasites are considered specific and sensitive, but often not applied in endemic areas of developing countries due to technical complexity. In the present study isothermal, nucleic acid sequence based amplification (NASBA) was coupled to oligochromatography (OC) to develop a simplified detection method for the diagnosis of leishmaniasis. NASBA-OC, detecting Leishmania RNA, was evaluated using clinical samples from visceral leishmaniasis patients from East Africa (n = 30) and cutaneous leishmaniasis from South America (n = 70) and appropriate control samples.


RESULTS
Analytical sensitivity was 10 parasites/ml of spiked blood, and 1 parasite/ml of culture. Diagnostic sensitivity of NASBA-OC was 93.3% (95% CI: 76.5%-98.8%) and specificity was 100% (95% CI: 91.1%-100%) on blood samples, while sensitivity and specificity on skin biopsy samples was 98.6% (95% CI: 91.2%-99.9%) and 100% (95% CI: 46.3%-100%), respectively.


CONCLUSIONS
The NASBA-OC format brings implementation of molecular diagnosis of leishmaniasis in resource poor countries one step closer.
Mugasa
Claire M
CM
Koninklijk Instituut voor de Tropen (KIT)/Royal Tropical Institute, KIT Biomedical Research, Amsterdam, the Netherlands. h.schallig@kit.nl.
Laurent
Thierry
T
Schoone
Gerard J
GJ
Basiye
Frank L
FL
Saad
Alfarazdeg A
AA
El Safi
Sayda
S
Kager
Piet A
PA
Schallig
Henk Dfh
HD