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Publication Details
AFRICAN RESEARCH NEXUS
SHINING A SPOTLIGHT ON AFRICAN RESEARCH
immunology and microbiology
An approach for interlaboratory comparison of conventional and real-time PCR assays for diagnosis of human leishmaniasis
Experimental Parasitology, Volume 134, No. 3, Year 2013
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Description
Protozoa of the Leishmania genus are transmitted to humans by the bite of infected sandflies, and are the causative agents of leishmaniasis which ranges from cutaneous to visceral clinical forms. The definitive diagnosis of leishmaniasis has relied traditionally on parasite demonstration, either by microscopy or culture; in the last years, diagnosis based on PCR methods has overcome some drawbacks of traditional methods, increasing sensitivity and allowing using less invasive sampling for diagnosis. However, there are not defined protocols and almost each laboratory applies its own in-house method. Although there are several studies comparing the performance of different methods within the same laboratory, those addressing interlaboratory comparison are scarce, in spite of the growing number of collaborative projects between partners from different leishmaniasis endemic and non-endemic countries. In this work we propose a protocol for interlaboratory comparison of conventional and real-time PCR methods involving four participant laboratories from four different endemic regions in four continents; the protocol includes a quality control step and reduces the variability among the samples tested by each participant. A panel of 77 samples from human origin and 9 from different parasite strains was blindly tested by the participants, aiming to assess the sensitivity of the different methods as well as their usefulness for species identification. Real-time PCR methods targeting the kDNA minicircles returned the highest sensitivity, while both PCR targeting ITS-1 and further HaeIII digestion and a combined algorithm including hsp70 PCR and restriction fragment length polymorphism analysis were the most appropriate approaches for species identification. © 2013 Elsevier Inc.
Authors & Co-Authors
Cruz, Israel
Spain, Majadahonda
Centro Nacional de Microbiologia
Millet, Aurélie
Tunisia, Tunis
Institut Pasteur de Tunis
Carrillo, E.
Spain, Majadahonda
Centro Nacional de Microbiologia
Chenik, Mehdi
Tunisia, Tunis
Institut Pasteur de Tunis
Salotra, Poonam
India, New Delhi
National Institute of Pathology
Verma, Sandeep
India, New Delhi
National Institute of Pathology
Veland, Nicolas
Peru, Lima
Universidad Peruana Cayetano Heredia, Instituto de Medicina Tropical Alexander Von Humboldt
Jara, Marlene
Peru, Lima
Universidad Peruana Cayetano Heredia, Instituto de Medicina Tropical Alexander Von Humboldt
Adaui, Vanessa
Peru, Lima
Universidad Peruana Cayetano Heredia, Instituto de Medicina Tropical Alexander Von Humboldt
Castrillón, Carlos
Peru, Lima
Universidad Peruana Cayetano Heredia, Instituto de Medicina Tropical Alexander Von Humboldt
Arevalo, Jorge Luis
Peru, Lima
Universidad Peruana Cayetano Heredia, Instituto de Medicina Tropical Alexander Von Humboldt
Moreno, Javier
Spain, Majadahonda
Centro Nacional de Microbiologia
Cañavate, Carmen
Spain, Majadahonda
Centro Nacional de Microbiologia
Statistics
Citations: 66
Authors: 13
Affiliations: 4
Identifiers
Doi:
10.1016/j.exppara.2013.03.026
ISSN:
00144894
e-ISSN:
10902449