PCR-RLFP characterization of Leishmania spp. in domestic animals from the south-western border of Brazil
Abstract The aim of this study was to characterize Leishmania spp. from canine and feline samples using Polymerase Chain Reaction (PCR)- Restriction Fragment Length Polymorphism (RFLP). It was conducted in the southern region of Brazil, located at border crossings to Argentina and Uruguay. Samples w...
Published in: | Revista Brasileira de Parasitologia Veterinária |
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Colégio Brasileiro de Parasitologia Veterinaria
2022
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ftdoajarticles:oai:doaj.org/article:19edc7c75c164bb2958db79ea888481f 2023-05-15T15:50:33+02:00 PCR-RLFP characterization of Leishmania spp. in domestic animals from the south-western border of Brazil Gabriela Döwich Pradella Taiane Acunha Escobar Thália Pacheco dos Santos Rammy Campos Vargas Geórgia Camargo Góss Patricia Aline Gröhs Ferrareze Lívia Kmetzsch Rosa e Silva Luísa Zuravski Karina Braccini Pereira Claudia Acosta Duarte Irina Lübeck 2022-07-01T00:00:00Z https://doi.org/10.1590/s1984-29612022035 https://doaj.org/article/19edc7c75c164bb2958db79ea888481f EN ES PT eng spa por Colégio Brasileiro de Parasitologia Veterinaria http://www.scielo.br/scielo.php?script=sci_arttext&pid=S1984-29612022000300301&lng=en&tlng=en http://www.scielo.br/pdf/rbpv/v31n3/1984-2961-rbpv-31-3-e005222.pdf https://doaj.org/toc/1984-2961 1984-2961 doi:10.1590/s1984-29612022035 https://doaj.org/article/19edc7c75c164bb2958db79ea888481f Revista Brasileira de Parasitologia Veterinária, Vol 31, Iss 3 (2022) Canine diagnosis feline Leishmaniasis molecular biology Animal culture SF1-1100 article 2022 ftdoajarticles https://doi.org/10.1590/s1984-29612022035 2022-12-30T21:15:16Z Abstract The aim of this study was to characterize Leishmania spp. from canine and feline samples using Polymerase Chain Reaction (PCR)- Restriction Fragment Length Polymorphism (RFLP). It was conducted in the southern region of Brazil, located at border crossings to Argentina and Uruguay. Samples were collected from 116 dogs (Canis lupus familiaris) and 89 cats (Felis catus). The PCR was performed to screen for an LT1 fragment from kinetoplast DNA (kDNA) target gene, and positive samples were subjected to a second PCR for an internal transcribed spacers (ITS1) region from ribosomal DNA (rDNA) target. RFLP was performed using the Haemophilus aegyptius (HAE III) restriction endonuclease (Fermentas ®). Positive samples by PCR ITS1 were sequenced and deposited in NCBI GenBank, and a phylogenetic analysis was developed. We found that 12.9% (15/116) of the samples from dogs were positive. All the 89 cat samples were negative. Positive samples were tested against Leishmania reference strains presenting different patterns in PCR-RFLP, and these samples showed bands denoting similarity to the standard species of Leishmania infantum, proven through sequencing and phylogenetic analysis. The RFLP technique, alone, was shown to be feasible for practical application and confirmation of the involved Leishmania spp. Article in Journal/Newspaper Canis lupus Directory of Open Access Journals: DOAJ Articles Argentina Uruguay Revista Brasileira de Parasitologia Veterinária 31 3 |
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collection |
Directory of Open Access Journals: DOAJ Articles |
op_collection_id |
ftdoajarticles |
language |
English Spanish Portuguese |
topic |
Canine diagnosis feline Leishmaniasis molecular biology Animal culture SF1-1100 |
spellingShingle |
Canine diagnosis feline Leishmaniasis molecular biology Animal culture SF1-1100 Gabriela Döwich Pradella Taiane Acunha Escobar Thália Pacheco dos Santos Rammy Campos Vargas Geórgia Camargo Góss Patricia Aline Gröhs Ferrareze Lívia Kmetzsch Rosa e Silva Luísa Zuravski Karina Braccini Pereira Claudia Acosta Duarte Irina Lübeck PCR-RLFP characterization of Leishmania spp. in domestic animals from the south-western border of Brazil |
topic_facet |
Canine diagnosis feline Leishmaniasis molecular biology Animal culture SF1-1100 |
description |
Abstract The aim of this study was to characterize Leishmania spp. from canine and feline samples using Polymerase Chain Reaction (PCR)- Restriction Fragment Length Polymorphism (RFLP). It was conducted in the southern region of Brazil, located at border crossings to Argentina and Uruguay. Samples were collected from 116 dogs (Canis lupus familiaris) and 89 cats (Felis catus). The PCR was performed to screen for an LT1 fragment from kinetoplast DNA (kDNA) target gene, and positive samples were subjected to a second PCR for an internal transcribed spacers (ITS1) region from ribosomal DNA (rDNA) target. RFLP was performed using the Haemophilus aegyptius (HAE III) restriction endonuclease (Fermentas ®). Positive samples by PCR ITS1 were sequenced and deposited in NCBI GenBank, and a phylogenetic analysis was developed. We found that 12.9% (15/116) of the samples from dogs were positive. All the 89 cat samples were negative. Positive samples were tested against Leishmania reference strains presenting different patterns in PCR-RFLP, and these samples showed bands denoting similarity to the standard species of Leishmania infantum, proven through sequencing and phylogenetic analysis. The RFLP technique, alone, was shown to be feasible for practical application and confirmation of the involved Leishmania spp. |
format |
Article in Journal/Newspaper |
author |
Gabriela Döwich Pradella Taiane Acunha Escobar Thália Pacheco dos Santos Rammy Campos Vargas Geórgia Camargo Góss Patricia Aline Gröhs Ferrareze Lívia Kmetzsch Rosa e Silva Luísa Zuravski Karina Braccini Pereira Claudia Acosta Duarte Irina Lübeck |
author_facet |
Gabriela Döwich Pradella Taiane Acunha Escobar Thália Pacheco dos Santos Rammy Campos Vargas Geórgia Camargo Góss Patricia Aline Gröhs Ferrareze Lívia Kmetzsch Rosa e Silva Luísa Zuravski Karina Braccini Pereira Claudia Acosta Duarte Irina Lübeck |
author_sort |
Gabriela Döwich Pradella |
title |
PCR-RLFP characterization of Leishmania spp. in domestic animals from the south-western border of Brazil |
title_short |
PCR-RLFP characterization of Leishmania spp. in domestic animals from the south-western border of Brazil |
title_full |
PCR-RLFP characterization of Leishmania spp. in domestic animals from the south-western border of Brazil |
title_fullStr |
PCR-RLFP characterization of Leishmania spp. in domestic animals from the south-western border of Brazil |
title_full_unstemmed |
PCR-RLFP characterization of Leishmania spp. in domestic animals from the south-western border of Brazil |
title_sort |
pcr-rlfp characterization of leishmania spp. in domestic animals from the south-western border of brazil |
publisher |
Colégio Brasileiro de Parasitologia Veterinaria |
publishDate |
2022 |
url |
https://doi.org/10.1590/s1984-29612022035 https://doaj.org/article/19edc7c75c164bb2958db79ea888481f |
geographic |
Argentina Uruguay |
geographic_facet |
Argentina Uruguay |
genre |
Canis lupus |
genre_facet |
Canis lupus |
op_source |
Revista Brasileira de Parasitologia Veterinária, Vol 31, Iss 3 (2022) |
op_relation |
http://www.scielo.br/scielo.php?script=sci_arttext&pid=S1984-29612022000300301&lng=en&tlng=en http://www.scielo.br/pdf/rbpv/v31n3/1984-2961-rbpv-31-3-e005222.pdf https://doaj.org/toc/1984-2961 1984-2961 doi:10.1590/s1984-29612022035 https://doaj.org/article/19edc7c75c164bb2958db79ea888481f |
op_doi |
https://doi.org/10.1590/s1984-29612022035 |
container_title |
Revista Brasileira de Parasitologia Veterinária |
container_volume |
31 |
container_issue |
3 |
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1766385527392567296 |