Article
An Innovative Field-Applicable Molecular Test to Diagnose Cutaneous Leishmania Viannia spp. Infections
Registro en:
SAIDARRIAGA, Omar A. et al. An Innovative Field-Applicable Molecular Test to Diagnose Cutaneous Leishmania Viannia spp. Infections. PLoS Negl Trop Dis , v.10, n.4, e0004638, 12p, Apr. 2016.
1932-6203
10.1371/journal. pntd.0004638
Autor
Saldarriaga, Omar A.
Castellanos-Gonzalez, Alejandro
Porrozzi, Renato
Baldeviano, Gerald C.
Lescano, Andrés G.
Los Santos, Maxy B. de
Fernandez, Olga L.
Saravia, Nancy G.
Costa, Erika
Melby, Peter C.
Travi, Bruno L.
Resumen
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.