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Molecular Identification of Parasites Causing Cutaneous Leishmaniasis in Panama

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  • 1 Instituto Conmemorativo Gorgas de Estudios de la Salud (ICGES), Panama, Panama;
  • 2 Facultad de Ciencias Naturales, Exactas y Tecnología, Universidad de Panamá, Panama, Panama;
  • 3 Facultad de Medicina Veterinaria, Universidad de Panamá, Panama, Panama;
  • 4 Walter Reed Army Institute of Research, Silver Spring, Maryland;
  • 5 U.S. Naval Medical Research Unit No. 6, Lima, Peru;
  • 6 Centro de Investigación y Diagnóstico de Enfermedades Parasitarias (CIDEP), Facultad de Medicina, Universidad de Panamá, Panama, Panama

ABSTRACT

Isolates from 475 cutaneous leishmaniasis (CL) patients from three endemic regions were studied by three typing techniques. The molecular analysis from lesion scrapings based on hsp70 PCR-restriction fragment length polymorphism (RFLP) showed that 78.1% (371/475) restriction patterns corresponded to Leishmania (Viannia) panamensis, 19% (90/475) to Leishmania (Viannia) guyanensis, and 3.0% (14/475) to Leishmania (Viannia) braziliensis. Promastigotes isolated by culture from lesions of 228 patients (48.0%, 228/475) were identified by multi-locus enzyme electrophoresis. Of them, 95.2% (217/228) were typified as L. (V.) panamensis, 1.3% (3/228) as L. (V.) guyanensis, 2.2% (5/228) as L. (V.) braziliensis, and 1.3% (3/228) as hybrids (L. [V.] braziliensis/L. [V.] panamensis). However, a partial sequencing analysis of the hsp70 gene from 77 selected samples showed 16.9% (13/77) typified as L. (V.) panamensis, 68.8% (53/77) as Leishmania (V.) sp., 1, 3.9% (3/77) as L. (V.) guyanensis, 1.3% (1/77) as L. (V.) braziliensis outlier, 2.6% (2/77) as Leishmania (Viannia) naiffi, 2.6% as (2/77) Leishmania (V.) sp., and 2 and 3.9% (3/77) hybrid isolates of L. (V.) braziliensis/L. (V.) guyanensis. These results confirm L. (V.) panamensis as the predominant species and cause of CL lesions in Panama and that L. (V.) guyanensis, L. (V.) braziliensis, and L. (V.) naiffi are circulating to a lower degree. Furthermore, the determination of parasite isolates belonging to atypical clusters and hybrid isolates suggests the circulation of genetic variants with important implications for the epidemiology and clinical follow-up of CL in Panama. No evidence of the existence of parasites of the Leishmania (Leishmania) mexicana complex in Panamanian territory was found in this study.

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Author Notes

Address correspondence to Néstor Sosa or Azael Saldaña, Instituto Conmemorativo Gorgas de Estudios de la Salud, Avenida Justo Arosemena, Calle 35, Calidonia, Panama 0816-02593, Panamá. E-mails: nsosa@gorgas.gob.pa or asaldana@gorgas.gob.pa

Financial support: This study was funded by the U.S. Army Medical Materiel Development Activity (USAMMDA), U.S. Army Medical Research and Materiel Command, and by the Instituto Conmemorativo Gorgas de Estudios de la Salud (ICGES), Panamá.

Disclosure: The investigators have adhered to the policies for protection of human subjects as prescribed in Army Regulation 70–25.

Authors’ addresses: Aracelis del C. Miranda, Kadir A. González, Franklyn Samudio, Vanessa J. Pineda, José E. Calzada, and Azael Saldaña, Departamento de Investigaciones en Parasitología, Instituto Conmemorativo Gorgas de Estudios de la Salud (ICGES), Panama, Panama, E-mails: ara04amp@yahoo.com, kgonzalez@gorgas.gob.pa, fsamudio@gorgas.gob.pa, vpineda@gorgas.gob.pa, jcalzada@gorgas.gob.pa, and asaldana@gorgas.gob.pa. Zeuz Capitan-Barrios, José A. Suárez, Betsi Ortiz, Juan M. Pascale, and Néstor Sosa, Unidad de Diagnóstico, Investigación Clínica y Medicina Tropical, Instituto Conmemorativo Gorgas de Estudios de la Salud (ICGES), Panama, Panama, E-mails: zcapitan@gmail.com, jsuarez@gorgas.gob.pa, bortiz@gorgas.gob.pa, jpascale@gorgas.gob.pa, and drnsosa@gmail.con. Ana Jiménez, Unidad de Diagnóstico, Investigación Clínica y Medicina Tropical, Instituto Conmemorativo Gorgas de Estudios de la Salud (ICGES), Panama, Panama, and EAP Marc Aureli, ICS Barcelona Ciutat, Barcelona, Spain, E-mail: anajimenezlozano@gmail.com. Juan Castillo and Yaxelis Mendoza, Departamento de Investigaciones en Genómica y Proteómica, Instituto Conmemorativo Gorgas de Estudios de la Salud (ICGES), Panama, Panama, E-mails: jcastillo@gorgas.gob.pa and ymendoza@gorgas.gob.pa. Juan Méndez, Walter Reed Army Institute of Research, Silver Spring, MD, E-mail: jmendez2808@comcast.net. Max Grögl, US Naval Medical Research Unit No. 6, in Lima, Peru, E-mail: max.grogl@gmail.com.

These authors contributed equally to this work.

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