New Primers for Detection and Differentiation between Leishmania viannia and L. leishmania Subgenera by Polymerase Chain Reaction
Background: Leishmania is the parasitic protozoan responsible for leishmaniases, a disease that can cause a range of cutaneous, mucosal, and visceral infections. Two subgenera L. Viannia and L. Leishmania are known to infect humans in the tropics and subtropics of the Americas. The aim of the prese...
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| Format: | Article |
| Language: | English |
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Tehran University of Medical Sciences
2023-09-01
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| Series: | Iranian Journal of Parasitology |
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| Online Access: | https://ijpa.tums.ac.ir/index.php/ijpa/article/view/3513 |
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| author | Manuel Calvopina David Fonseca-Carrera Irina Villacrés-Granda Alberto Toapanta Carlos Chiluisa-Guacho Carlos Bastidas-Caldes |
| author_facet | Manuel Calvopina David Fonseca-Carrera Irina Villacrés-Granda Alberto Toapanta Carlos Chiluisa-Guacho Carlos Bastidas-Caldes |
| author_sort | Manuel Calvopina |
| collection | DOAJ |
| description |
Background: Leishmania is the parasitic protozoan responsible for leishmaniases, a disease that can cause a range of cutaneous, mucosal, and visceral infections. Two subgenera L. Viannia and L. Leishmania are known to infect humans in the tropics and subtropics of the Americas. The aim of the present study was to develop a new pair of primers for the two subgenera and test in clinical samples.
Methods: We designed two new pairs of primers for a PCR method from two conserved genes, cysteine proteinase B (cpb) and N-acetylglucosamine-6-phosfate deacetylase-like protein (nagA), as specific markers for those two respective subgenera. Primers were tested with 16 microscopical positive clinical samples from the Amazon region of Ecuador obtained in 2010-2020 period.
Results: The cpb presented a band of 172 bp and the nagA a band of 300 bp, thus clearly differentiating L. viannia from L. leishmania. Additionally, primers identified and differentiated the clinical samples in the two subgenera.
Conclusion: The new primers targeting different two genes and standardized in a PCR assay could identified and differentiated Leishmania parasites at subgenus level. This protocol could be used for Leishmania genus identification and diagnosis at the subgenus level and for determining the parasite's geographical distribution where different Leishmania subgenera are found in the same area.
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| first_indexed | 2024-03-11T22:05:48Z |
| format | Article |
| id | doaj.art-4f713e79a8fd4a9faebea0e4da7339a9 |
| institution | Directory Open Access Journal |
| issn | 1735-7020 2008-238X |
| language | English |
| last_indexed | 2024-03-11T22:05:48Z |
| publishDate | 2023-09-01 |
| publisher | Tehran University of Medical Sciences |
| record_format | Article |
| series | Iranian Journal of Parasitology |
| spelling | doaj.art-4f713e79a8fd4a9faebea0e4da7339a92023-09-25T05:54:35ZengTehran University of Medical SciencesIranian Journal of Parasitology1735-70202008-238X2023-09-01183New Primers for Detection and Differentiation between Leishmania viannia and L. leishmania Subgenera by Polymerase Chain ReactionManuel Calvopina0David Fonseca-Carrera1Irina Villacrés-Granda2Alberto Toapanta3Carlos Chiluisa-Guacho4Carlos Bastidas-Caldes5One Health Research Group, Faculty of Medicine, Universidad de las Américas (UDLA), Quito, EcuadorBiotechnology Engineering, Faculty of Engineering and Applied Sciences (FICA), Universidad de las Américas (UDLA), Quito, EcuadorBiotechnology Engineering, Faculty of Engineering and Applied Sciences (FICA), Universidad de las Américas (UDLA), Quito, EcuadorBiotechnology Engineering, Faculty of Engineering and Applied Sciences (FICA), Universidad de las Américas (UDLA), Quito, EcuadorNational Institute for Investigation in Public Health “Leopoldo Izquieta Pérez (INSPI)”, Tena, EcuadorDoctoral Program in Public and Animal Health, Universidad de Extremadura, Extremadura, Spain Background: Leishmania is the parasitic protozoan responsible for leishmaniases, a disease that can cause a range of cutaneous, mucosal, and visceral infections. Two subgenera L. Viannia and L. Leishmania are known to infect humans in the tropics and subtropics of the Americas. The aim of the present study was to develop a new pair of primers for the two subgenera and test in clinical samples. Methods: We designed two new pairs of primers for a PCR method from two conserved genes, cysteine proteinase B (cpb) and N-acetylglucosamine-6-phosfate deacetylase-like protein (nagA), as specific markers for those two respective subgenera. Primers were tested with 16 microscopical positive clinical samples from the Amazon region of Ecuador obtained in 2010-2020 period. Results: The cpb presented a band of 172 bp and the nagA a band of 300 bp, thus clearly differentiating L. viannia from L. leishmania. Additionally, primers identified and differentiated the clinical samples in the two subgenera. Conclusion: The new primers targeting different two genes and standardized in a PCR assay could identified and differentiated Leishmania parasites at subgenus level. This protocol could be used for Leishmania genus identification and diagnosis at the subgenus level and for determining the parasite's geographical distribution where different Leishmania subgenera are found in the same area. https://ijpa.tums.ac.ir/index.php/ijpa/article/view/3513Cpb geneDiagnosisLeishmania;Naga geneNew world;Primers |
| spellingShingle | Manuel Calvopina David Fonseca-Carrera Irina Villacrés-Granda Alberto Toapanta Carlos Chiluisa-Guacho Carlos Bastidas-Caldes New Primers for Detection and Differentiation between Leishmania viannia and L. leishmania Subgenera by Polymerase Chain Reaction Iranian Journal of Parasitology Cpb gene Diagnosis Leishmania; Naga gene New world; Primers |
| title | New Primers for Detection and Differentiation between Leishmania viannia and L. leishmania Subgenera by Polymerase Chain Reaction |
| title_full | New Primers for Detection and Differentiation between Leishmania viannia and L. leishmania Subgenera by Polymerase Chain Reaction |
| title_fullStr | New Primers for Detection and Differentiation between Leishmania viannia and L. leishmania Subgenera by Polymerase Chain Reaction |
| title_full_unstemmed | New Primers for Detection and Differentiation between Leishmania viannia and L. leishmania Subgenera by Polymerase Chain Reaction |
| title_short | New Primers for Detection and Differentiation between Leishmania viannia and L. leishmania Subgenera by Polymerase Chain Reaction |
| title_sort | new primers for detection and differentiation between leishmania viannia and l leishmania subgenera by polymerase chain reaction |
| topic | Cpb gene Diagnosis Leishmania; Naga gene New world; Primers |
| url | https://ijpa.tums.ac.ir/index.php/ijpa/article/view/3513 |
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