An Innovative Field-Applicable Molecular Test to Diagnose Cutaneous Leishmania Viannia spp. Infections.
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.
Division of Infectious Diseases, Department of Internal Medicine, University of Texas Medical Branch (UTMB), Galveston, Texas, United States of America.
Omar A
OA
Saldarriaga
Division of Infectious Diseases, Department of Internal Medicine, University of Texas Medical Branch (UTMB), Galveston, Texas, United States of America.|Center for Tropical Diseases (CTD), University of Texas Medical Branch (UTMB), Galveston, Texas, United States of America.
Alejandro
A
Castellanos-Gonzalez
Laboratório de Pesquisa em Leishmaniose, Instituto Oswaldo Cruz, Rio de Janeiro, Brazil.
Renato
R
Porrozzi
U.S. Naval Medical Research Unit No. 6 (NAMRU-6), Lima, Perú
Gerald C
GC
Baldeviano
U.S. Naval Medical Research Unit No. 6 (NAMRU-6), Lima, Perú|Facultad de Salud Pública y Administración, Universidad Peruana Cayetano Heredia, Lima, Peru.
Andrés G
AG
Lescano
U.S. Naval Medical Research Unit No. 6 (NAMRU-6), Lima, Perú
Maxy B
MB
de Los Santos
Centro Internacional de Entrenamiento e Investigaciones Médicas (CIDEIM), Cali, Colombia.
Olga L
OL
Fernandez
Centro Internacional de Entrenamiento e Investigaciones Médicas (CIDEIM), Cali, Colombia.
Nancy G
NG
Saravia
Laboratório de Pesquisa em Leishmaniose, Instituto Oswaldo Cruz, Rio de Janeiro, Brazil.
Erika
E
Costa
Division of Infectious Diseases, Department of Internal Medicine, University of Texas Medical Branch (UTMB), Galveston, Texas, United States of America.|Center for Tropical Diseases (CTD), University of Texas Medical Branch (UTMB), Galveston, Texas, United States of America.|Department of Microbiology and Immunology, University of Texas Medical Branch (UTMB), Galveston, Texas, United States of America.
Peter C
PC
Melby
Division of Infectious Diseases, Department of Internal Medicine, University of Texas Medical Branch (UTMB), Galveston, Texas, United States of America.|Center for Tropical Diseases (CTD), University of Texas Medical Branch (UTMB), Galveston, Texas, United States of America.|Department of Microbiology and Immunology, University of Texas Medical Branch (UTMB), Galveston, Texas, United States of America.
Bruno L
BL
Travi
D002846:Chromatography, Affinity; D017931:DNA Primers / Q000235:genetics; D018105:DNA, Kinetoplast / Q000235:genetics; D016054:DNA, Protozoan / Q000235:genetics; D006801:Humans; D007891:Leishmania / Q000235:genetics / Q000302:isolation & purification*; D016773:Leishmaniasis, Cutaneous / Q000175:diagnosis*; D025202:Molecular Diagnostic Techniques / Q000379:methods*; D021141:Nucleic Acid Amplification Techniques; D015345:Oligonucleotide Probes / Q000235:genetics; D019095:Point-of-Care Systems; D012680:Sensitivity and Specificity
D017931:DNA Primers; D018105:DNA, Kinetoplast; D016054:DNA, Protozoan; D015345:Oligonucleotide Probes