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https://www.arca.fiocruz.br/handle/icict/7204
DETECTION AND IDENTIFICATION OF LEISHMANIA SPECIES IN FIELD-CAPTURED PHLEBOTOMINE SANDFLIES BASED ON MINI-EXON GENE PCR
Author
Affilliation
Universidade de Sao Paulo. Instituto de Medicina Tropical de Sao Paulo. Sao Paulo, SP, Brazil/Universidade de Sao Paulo. Instituto de Ciencias Biomedicas. Sao Paulo, SP, Brazil
Fundação Oswaldo Cruz. Centro de Pesquisas Rene Rachou. Laboratorio de Entomologia Medica. Belo Horizonte, MG, Brazil
Secretaria de Estado da Saude. Laboratorio Regional de Entomologia. Dourados, MT, Brazil
Fundação Oswaldo Cruz. Centro de Pesquisas Rene Rachou. Laboratorio de Entomologia Medica. Belo Horizonte, MG, Brazil
Universidade de Sao Paulo. Faculdade de Saude Publica. Sao Paulo, SP, Brazil
Universidade de Sao Paulo. Instituto de Medicina Tropical de Sao Paulo. Sao Paulo, SP, Brazil/Universidade de Sao Paulo. Instituto de Ciencias Biomedicas. Sao Paulo, SP, Brazil
Universidade de Sao Paulo. Instituto de Medicina Tropical de Sao Paulo. Sao Paulo, SP, Brazil /Universidade de Sao Paulo. Departamento de Doenças Parasitárias e Infecciosas da Faculdade de Medicina. Sao Paulo, SP, Brazil
Fundação Oswaldo Cruz. Centro de Pesquisas Rene Rachou. Laboratorio de Entomologia Medica. Belo Horizonte, MG, Brazil
Secretaria de Estado da Saude. Laboratorio Regional de Entomologia. Dourados, MT, Brazil
Fundação Oswaldo Cruz. Centro de Pesquisas Rene Rachou. Laboratorio de Entomologia Medica. Belo Horizonte, MG, Brazil
Universidade de Sao Paulo. Faculdade de Saude Publica. Sao Paulo, SP, Brazil
Universidade de Sao Paulo. Instituto de Medicina Tropical de Sao Paulo. Sao Paulo, SP, Brazil/Universidade de Sao Paulo. Instituto de Ciencias Biomedicas. Sao Paulo, SP, Brazil
Universidade de Sao Paulo. Instituto de Medicina Tropical de Sao Paulo. Sao Paulo, SP, Brazil /Universidade de Sao Paulo. Departamento de Doenças Parasitárias e Infecciosas da Faculdade de Medicina. Sao Paulo, SP, Brazil
Abstract
Leishmaniasis is one of the most diverse and complex of all vector-borne diseases. Because it involves several overlapping species and sandfly vectors, the disease has a complex ecology and epidemiology. Adequate therapy and follow-up depend on parasitological diagnosis, but classical methods present several constraints when identifying species. We describe a polymerase chain reaction (PCR) which uses primers designed from mini-exon repetitive sequences that are specific for subgenus Leishmania Viannia (PV), as well as sequences with specificity for genus (PG) that can distinguish between Leishmania species from other insect flagellates with minor differences in PCR products. For standardization, these PCR were tested in experimentally infected sandflies, and Leishmania infection in these insects was successfully confirmed. This methodology identified a 3.9% infection rate in field-captured phlebotomine sandflies from an endemic region in Brazil. Natural infection by Leishmania species was identified in three samples of Lutzomyia longipalpis, of which two were Leishmania (L.) chagasi and one Leishmania (L.) amazonensis. Irrespective of specific epidemiological conclusions, the method used in this study was able to identify Leishmania infections both in experimentally infected and field-captured phlebotomine sandflies, and could be a useful tool in epidemiological studies and strategic planning for the control of human leishmaniasis.
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