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Publication Details
AFRICAN RESEARCH NEXUS
SHINING A SPOTLIGHT ON AFRICAN RESEARCH
medicine
Nucleic acid sequence-based amplification with oligochromatography for detection of Trypanosoma brucei in clinical samples
Journal of Clinical Microbiology, Volume 47, No. 3, Year 2009
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Description
Molecular tools, such as real-time nucleic acid sequence-based amplification (NASBA) and PCR, have been developed to detect Trypanosoma brucei parasites in blood for the diagnosis of human African trypanosomiasis (HAT). Despite good sensitivity, these techniques are not implemented in HAT control programs due to the high cost of the equipment, which is unaffordable for laboratories in developing countries where HAT is endemic. In this study, a simplified technique, oligochromatography (OC), was developed for the detection of amplification products of T. brucei 18S rRNA by NASBA. The T. brucei NASBA-OC test has analytical sensitivities of 1 to 10 parasites/ml on nucleic acids extracted from parasite culture and 10 parasites/ml on spiked blood. The test showed no reaction with nontarget pathogens or with blood from healthy controls. Compared to the composite standard applied in the present study, i.e., parasitological confirmation of a HAT case by direct microscopy or by microscopy after concentration of parasites using either a microhematocrit centrifugation technique or a mini-anion-exchange centrifugation technique, NASBA-OC on blood samples had a sensitivity of 73.0% (95% confidence interval, 60 to 83%), while standard expert microscopy had a sensitivity of 57.1% (95% confidence interval, 44 to 69%). On cerebrospinal fluid samples, NASBA-OC had a sensitivity of 88.2% (95% confidence interval, 75 to 95%) and standard microscopy had a sensitivity of 86.2% (95% confidence interval, 64 to 88%). The T. brucei NASBA-OC test developed in this study can be employed in field laboratories, because it does not require a thermocycler; a simple heat block or a water bath maintained at two different temperatures is sufficient for amplification. Copyright © 2009, American Society for Microbiology. All Rights Reserved.
Authors & Co-Authors
Mugasa, Claire Mack
Uganda, Kampala
Makerere University
Netherlands, Amsterdam
Royal Tropical Institute - Kit
Laurent, Thierry Cl
Netherlands, Amsterdam
Amsterdam Umc - University of Amsterdam
Schoone, Gerard J.
Netherlands, Amsterdam
Royal Tropical Institute - Kit
Kager, Petrus A.
Belgium, Gembloux
Coris Bioconcept
Lubega, George William
Uganda, Kampala
Makerere University
Schallig, H. D. F. H.
Netherlands, Amsterdam
Royal Tropical Institute - Kit
Statistics
Citations: 71
Authors: 6
Affiliations: 4
Identifiers
Doi:
10.1128/JCM.01430-08
ISSN:
00951137
Research Areas
Environmental