Ana M Montalvo1, Jorge Fraga1, Omaira Rodríguez2, Orestes Blanco1, Alejandro Llanos-Cuentas3, Ana L García4, Braulio M Valencia3, Carlos Muskus5, Gert Van der Auwera6, José M Requena7. 1. Departamento de Parasitología, Instituto de Medicina Tropical Pedro Kourí, La Habana, Cuba. 2. Laboratorio de referencia e investigación en enfermedades tropicales de sanidad militar, Bogotá, Colombia. 3. Instituto de Medicina Tropical Alexander von Humboldt, Universidad Peruana Cayetano Heredia, Lima, Perú 4. Universidad de San Simón, Cochabamba, Bolivia. 5. Programa de Estudio y Control de Enfermedades Tropicales, Universidad de Antioquia, Medellín, Colombia. 6. Biomedical Sciences Department, Institute of Tropical Medicine of Antwerp, Amberes, Bélgica. 7. Centro de Biología Molecular Severo Ochoa, Madrid, España.
Abstract
OBJECTIVES: Explore a new target for molecular diagnosis of Leishmania. MATERIALS AND METHODS: We evaluated the utility of the gene that encodes the heat shock protein 20-kDa (Hsp20) for detecting Leishmania by polymerase chain reaction (PCR). PCR was normalized and analytical parameters were determined, as well as the validity and diagnostic accuracy, and concordance with the PCR - 18S. PCR-Hsp20 with DNA was obtained from a group of clinical samples from different sources. RESULTS: The analytical parameters were adequate. The sensitivity obtained was 86% and the specificity was 100%. The concordance with the reference method was good (κ = 0.731), which supports its potential use for diagnosis. The possibility of subsequent identification of the species by sequencing the amplified product gives an additional advantage. CONCLUSIONS: The usefulness of this gene as a new target for the detection of Leishmania was demonstrated. Because of its potential, it is recommended to improve the sensitivity of the method and to evaluate it in different endemic regions.
OBJECTIVES: Explore a new target for molecular diagnosis of Leishmania. MATERIALS AND METHODS: We evaluated the utility of the gene that encodes the heat shock protein 20-kDa (Hsp20) for detecting Leishmania by polymerase chain reaction (PCR). PCR was normalized and analytical parameters were determined, as well as the validity and diagnostic accuracy, and concordance with the PCR - 18S. PCR-Hsp20 with DNA was obtained from a group of clinical samples from different sources. RESULTS: The analytical parameters were adequate. The sensitivity obtained was 86% and the specificity was 100%. The concordance with the reference method was good (κ = 0.731), which supports its potential use for diagnosis. The possibility of subsequent identification of the species by sequencing the amplified product gives an additional advantage. CONCLUSIONS: The usefulness of this gene as a new target for the detection of Leishmania was demonstrated. Because of its potential, it is recommended to improve the sensitivity of the method and to evaluate it in different endemic regions.