Universidad Peruana Cayetano Heredia

An approach for interlaboratory comparison of conventional and real-time PCR assays for diagnosis of human leishmaniasis

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dc.contributor.author Cruz, Israel
dc.contributor.author Millet, Aurélie
dc.contributor.author Carrillo, Eugenia
dc.contributor.author Chenik, Mehdi
dc.contributor.author Salotra, Poonam
dc.contributor.author Verma, Sandeep
dc.contributor.author Veland, Nicolás
dc.contributor.author Jara, Marlene
dc.contributor.author Adaui, Vanessa
dc.contributor.author Castrillón, Carlos
dc.contributor.author Arévalo Zelada, Jorge Luis
dc.contributor.author Moreno, Javier
dc.contributor.author Cañavate, Carmen
dc.date.accessioned 2022-01-04T20:29:58Z
dc.date.available 2022-01-04T20:29:58Z
dc.date.issued 2013
dc.identifier.uri https://hdl.handle.net/20.500.12866/10460
dc.description.abstract 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. en_US
dc.language.iso eng
dc.publisher Elsevier
dc.relation.ispartofseries Experimental Parasitology
dc.rights info:eu-repo/semantics/restrictedAccess
dc.rights.uri https://creativecommons.org/licenses/by-nc-nd/4.0/deed.es
dc.subject Leishmania en_US
dc.subject PCR en_US
dc.subject Diagnosis Interlaboratory en_US
dc.subject Comparison en_US
dc.title An approach for interlaboratory comparison of conventional and real-time PCR assays for diagnosis of human leishmaniasis en_US
dc.type info:eu-repo/semantics/article
dc.identifier.doi https://doi.org/10.1016/j.exppara.2013.03.026
dc.subject.ocde https://purl.org/pe-repo/ocde/ford#3.03.07
dc.relation.issn 1090-2449


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