4.5 Article

An Innovative Field-Applicable Molecular Test to Diagnose Cutaneous Leishmania Viannia spp. Infections

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PLOS NEGLECTED TROPICAL DISEASES
卷 10, 期 4, 页码 -

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PUBLIC LIBRARY SCIENCE
DOI: 10.1371/journal.pntd.0004638

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资金

  1. Armed Forces Health Surveillance Center, Global Emerging Infections Surveillance and Response System
  2. AFHSC/GEIS of the U.S. Department of Defense
  3. Congresionally Directed Medical Research Programs (CDMRP) Award [W81XWH-14-2-0196]
  4. Institute for Translational Sciences at the University of Texas Medical Branch (UTMB)
  5. Clinical and Translational Science Award from the National Center for Advancing Translational Sciences, National Institutes of Health [UL1 TR001439]
  6. MTT Award [UL1TR000071]
  7. Center for Tropical Diseases from UTMB
  8. Fogarty International Center of the US National Institutes of Health [2D43 TW007393]
  9. Departamento Administrativo de Ciencia, Tecnologia e Innovacion-COLCIENCIAS
  10. [PR130282]

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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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