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Free Content Accordance and concordance of PCR and NASBA followed by oligochromatography for the molecular diagnosis of Trypanosoma brucei and Leishmania

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Summary Objective  To evaluate the repeatability and reproducibility of four simplified molecular assays for the diagnosis of spp. or ssp. in a multicentre ring trial with seven participating laboratories. Methods  The tests are based on PCR or NASBA amplification of the parasites nucleic acids followed by rapid read-out by oligochromatographic dipstick (PCR-OC and NASBA-OC). Results  On purified nucleic acid specimens, the repeatability and reproducibility of the tests were -PRC-OC, 91.7% and 95.5%; -NASBA-OC, 95.8% and 100%; -PCR-OC, 95.9% and 98.1%; -NASBA-OC, 92.3% and 98.2%. On blood specimens spiked with parasites, the repeatability and reproducibility of the tests were -PRC-OC, 78.4% and 86.6%; -NASBA-OC, 81.5% and 89.0%; -PCR-OC, 87.1% and 91.7%; -NASBA-OC, 74.8% and 86.2%. Conclusion  As repeatability and reproducibility of the tests were satisfactory, further phase II and III evaluations in clinical and population specimens from disease endemic countries are justified.

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Objectif: 

Evaluer la répétabilité et la reproductibilité de quatre tests moléculaires simplifiés pour le diagnostic de Trypanosoma brucei spp. ou Leishmania ssp. dans un essai multicentrique avec 7 laboratoires participants. Méthodes: 

Les tests sont basés sur l’amplification par PCR ou NASBA des acides nucléiques des parasites suivie par la lecture rapide sur bandelette oligochromatographique (PCR-OC et NASBA-OC). Résultats: 

Sur les échantillons d’acide nucléique purifié, la répétabilité et la reproductibilité des tests ont été les suivantes: Tryp-PRC-OC, 91,7% et 95,5%; Tryp-NASBA-OC, 95,8% et 100%; Leish-PCR-OC, 95,9% et 98,1%; Leish-NASBA-OC, 92,3% et 98,2%. Sur les échantillons de sang mélangés avec des parasites la répétabilité et la reproductibilité des tests ont été les suivantes: Tryp-PRC-OC, 78,4% et 86,6%; Tryp-NASBA-OC, 81,5% et 89,0%; Leish-PCR-OC, 87,1% et 91,7%; Leish-NASBA-OC, 74,8% et 86,2%. Conclusion: 

As repeatability and reproducibility of the tests were satisfactory, further phase II and III evaluations in clinical and population specimens from disease endemic countries are justified. Conclusion: 

Comme la répétabilité et la reproductibilité des tests étaient satisfaisantes, des évaluations ultérieures de phase II et III sur des échantillons cliniques provenance de populations des pays endémiques sont justifiées.
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Keywords: Leishmaniasis; NASBA; PCR; diagnostic; diagnostics; diagnóstico; leishmaniasis; leishmaniose; nucleic acid sequence–based amplification; oligochromatographie; oligochromatography; oligocromatografía; polymerase chain reaction; tripanosomiasis; trypanosomiais; trypanosomiase

Document Type: Research Article

Affiliations: 1:  Department of Parasitology, Institute of Tropical Medicine, Antwerp, Belgium 2:  Biomedical Research, Royal Tropical Institute, Amsterdam, The Netherlands 3:  Coris BioConcept, Gembloux, Belgium 4:  Department of Parasitology, Institut National de Recherche Biomédicale, Kinshasa, Congo 5:  Medical Faculty, University of Khartoum, Khartoum, Sudan 6:  Kenya Medical Research Centre, Nairobi, Kenya 7:  Department of Veterinary Parasitology and Microbiology, Makerere University, Kampala, Uganda 8:  Academic Medical Centre, Division of Infectious Diseases, Tropical Medicine and AIDS, Amsterdam, The Netherlands

Publication date: 2010-07-01

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