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AN INNOVATIVE FIELD-APPLICABLE MOLECULAR TEST TO DIAGNOSE CUTANEOUS LEISHMANIA VIANNIA SPP. INFECTIONS
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University of Texas Medical Branch (UTMB). Department of Internal Medicine. Division of Infectious Diseases. Galveston, Texas, USA.
University of Texas Medical Branch (UTMB). Department of Internal Medicine. Division of Infectious Diseases. Galveston, Texas, USA / University of Texas Medical Branch (UTMB),. Center for Tropical Diseases (CTD). Galveston, Texas, USA.
Fundação Oswaldo Cruz. Instituto Oswaldo Cruz. Laboratório de Pesquisa em Leishmaniose. Rio de Janeiro, RJ, Brasil.
U.S. Naval Medical Research Unit No. 6 (NAMRU-6). Lima, Peru.
U.S. Naval Medical Research Unit No. 6 (NAMRU-6). Lima, Peru / Universidad Peruana Cayetano Heredia. Lima, Peru.
U.S. Naval Medical Research Unit No. 6 (NAMRU-6). Lima, Peru.
Centro Internacional de Entrenamiento e Investigaciones Médicas (CIDEIM). Cali, Colombia.
Centro Internacional de Entrenamiento e Investigaciones Médicas (CIDEIM). Cali, Colombia.
Fundação Oswaldo Cruz. Instituto Oswaldo Cruz. Laboratório de Pesquisa em Leishmaniose. Rio de Janeiro, RJ. Brasil.
University of Texas Medical Branch (UTMB). Department of Internal Medicine. Division of Infectious Diseases. Galveston, Texas, USA / University of Texas Medical Branch (UTMB),. Center for Tropical Diseases (CTD). Galveston, Texas, USA / University of Texas Medical Branch (UTMB). Department of Microbiology and Immunology. Galveston, Texas, USA.
University of Texas Medical Branch (UTMB). Department of Internal Medicine. Division of Infectious Diseases. Galveston, Texas, USA / University of Texas Medical Branch (UTMB),. Center for Tropical Diseases (CTD). Galveston, Texas, USA / University of Texas Medical Branch (UTMB). Department of Microbiology and Immunology. Galveston, Texas, USA.
University of Texas Medical Branch (UTMB). Department of Internal Medicine. Division of Infectious Diseases. Galveston, Texas, USA / University of Texas Medical Branch (UTMB),. Center for Tropical Diseases (CTD). Galveston, Texas, USA.
Fundação Oswaldo Cruz. Instituto Oswaldo Cruz. Laboratório de Pesquisa em Leishmaniose. Rio de Janeiro, RJ, Brasil.
U.S. Naval Medical Research Unit No. 6 (NAMRU-6). Lima, Peru.
U.S. Naval Medical Research Unit No. 6 (NAMRU-6). Lima, Peru / Universidad Peruana Cayetano Heredia. Lima, Peru.
U.S. Naval Medical Research Unit No. 6 (NAMRU-6). Lima, Peru.
Centro Internacional de Entrenamiento e Investigaciones Médicas (CIDEIM). Cali, Colombia.
Centro Internacional de Entrenamiento e Investigaciones Médicas (CIDEIM). Cali, Colombia.
Fundação Oswaldo Cruz. Instituto Oswaldo Cruz. Laboratório de Pesquisa em Leishmaniose. Rio de Janeiro, RJ. Brasil.
University of Texas Medical Branch (UTMB). Department of Internal Medicine. Division of Infectious Diseases. Galveston, Texas, USA / University of Texas Medical Branch (UTMB),. Center for Tropical Diseases (CTD). Galveston, Texas, USA / University of Texas Medical Branch (UTMB). Department of Microbiology and Immunology. Galveston, Texas, USA.
University of Texas Medical Branch (UTMB). Department of Internal Medicine. Division of Infectious Diseases. Galveston, Texas, USA / University of Texas Medical Branch (UTMB),. Center for Tropical Diseases (CTD). Galveston, Texas, USA / University of Texas Medical Branch (UTMB). Department of Microbiology and Immunology. Galveston, Texas, USA.
Abstract
Cutaneous and mucosal leishmaniasis is widely distributed in Central and South America.
Leishmania of the Viannia subgenus are the most frequent species infecting humans. L. (V.)
braziliensis, L. (V.) panamensis are also responsible for metastatic mucosal leishmaniasis.
Conventional or real time PCR is a more sensitive diagnostic test than microscopy, but the
cost and requirement for infrastructure and trained personnel makes it impractical in most
endemic regions. Primary health systems need a sensitive and specific point of care (POC)
diagnostic tool. We developed a novel POC molecular diagnostic test for cutaneous leishmaniasis
caused by Leishmania (Viannia) spp. Parasite DNA was amplified using isothermal
Recombinase Polymerase Amplification (RPA) with primers and probes that targeted the
kinetoplast DNA. The amplification product was detected by naked eye with a lateral flow
(LF) immunochromatographic strip. The RPA-LF had an analytical sensitivity equivalent to
0.1 parasites per reaction. The test amplified the principal L. Viannia species from multiple
countries: L. (V.) braziliensis (n = 33), L. (V.) guyanensis (n = 17), L. (V.) panamensis (n = 9).
The less common L. (V.) lainsoni, L. (V.) shawi, and L. (V.) naiffi were also amplified. No
amplification was observed in parasites of the L. (Leishmania) subgenus. In a small number
of clinical samples (n = 13) we found 100% agreement between PCR and RPA-LF. The
high analytical sensitivity and clinical validation indicate the test could improve the efficiency
of diagnosis, especially in chronic lesions with submicroscopic parasite burdens. Field
implementation of the RPA-LF test could contribute to management and control of cutaneous
and mucosal leishmaniasis.
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