PERFORMANCE OF CONVENTIONAL PCRs BASED ON PRIMERS DIRECTED TO NUCLEAR AND MITOCHONDRIAL GENES FOR THE DETECTION AND IDENTIFICATION OF Leishmania spp.
In visceral leishmaniasis, the detection of the agent is of paramount importance to identify reservoirs of infection. Here, we evaluated the diagnostic attributes of PCRs based on primers directed to cytochrome-B (cytB), cytochrome-oxidase-subunit II (coxII), cytochrome-C (cytC), and the minicircle-...
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Universidade de São Paulo (USP)
2016
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ftdoajarticles:oai:doaj.org/article:a2a8de8d60334f96971bd5c381d3105c 2024-09-09T19:27:04+00:00 PERFORMANCE OF CONVENTIONAL PCRs BASED ON PRIMERS DIRECTED TO NUCLEAR AND MITOCHONDRIAL GENES FOR THE DETECTION AND IDENTIFICATION OF Leishmania spp. Estela Gallucci LOPES Carlos Alberto GERALDO JUNIOR Arlei MARCILI Ricardo Duarte SILVA Lara Borges KEID Trícia Maria Ferreira da Silva OLIVEIRA Rodrigo Martins SOARES 2016-01-01T00:00:00Z https://doi.org/10.1590/S1678-9946201658041 https://doaj.org/article/a2a8de8d60334f96971bd5c381d3105c EN eng Universidade de São Paulo (USP) http://www.scielo.br/scielo.php?script=sci_arttext&pid=S0036-46652016005000231&lng=en&tlng=en https://doaj.org/toc/1678-9946 1678-9946 doi:10.1590/S1678-9946201658041 https://doaj.org/article/a2a8de8d60334f96971bd5c381d3105c Revista do Instituto de Medicina Tropical de São Paulo, Vol 58, Iss 0 (2016) Leishmania spp kDNA Mitochondrial genes PCR Phylogeny Dogs Arctic medicine. Tropical medicine RC955-962 Infectious and parasitic diseases RC109-216 article 2016 ftdoajarticles https://doi.org/10.1590/S1678-9946201658041 2024-08-05T17:49:30Z In visceral leishmaniasis, the detection of the agent is of paramount importance to identify reservoirs of infection. Here, we evaluated the diagnostic attributes of PCRs based on primers directed to cytochrome-B (cytB), cytochrome-oxidase-subunit II (coxII), cytochrome-C (cytC), and the minicircle-kDNA. Although PCRs directed to cytB, coxII, cytC were able to detect different species of Leishmania, and the nucleotide sequence of their amplicons allowed the unequivocal differentiation of species, the analytical and diagnostic sensitivity of these PCRs were much lower than the analytical and diagnostic sensitivity of the kDNA-PCR. Among the 73 seropositive animals, the asymptomatic dogs had spleen and bone marrow samples collected and tested; only two animals were positive by PCRs based on cytB, coxII, and cytC, whereas 18 were positive by the kDNA-PCR. Considering the kDNA-PCR results, six dogs had positive spleen and bone marrow samples, eight dogs had positive bone marrow results but negative results in spleen samples and, in four dogs, the reverse situation occurred. We concluded that PCRs based on cytB, coxII, and cytC can be useful tools to identify Leishmania species when used in combination with automated sequencing. The discordance between the results of the kDNA-PCR in bone marrow and spleen samples may indicate that conventional PCR lacks sensitivity for the detection of infected dogs. Thus, primers based on the kDNA should be preferred for the screening of infected dogs. Article in Journal/Newspaper Arctic Directory of Open Access Journals: DOAJ Articles Arctic Revista do Instituto de Medicina Tropical de São Paulo 58 0 |
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topic |
Leishmania spp kDNA Mitochondrial genes PCR Phylogeny Dogs Arctic medicine. Tropical medicine RC955-962 Infectious and parasitic diseases RC109-216 |
spellingShingle |
Leishmania spp kDNA Mitochondrial genes PCR Phylogeny Dogs Arctic medicine. Tropical medicine RC955-962 Infectious and parasitic diseases RC109-216 Estela Gallucci LOPES Carlos Alberto GERALDO JUNIOR Arlei MARCILI Ricardo Duarte SILVA Lara Borges KEID Trícia Maria Ferreira da Silva OLIVEIRA Rodrigo Martins SOARES PERFORMANCE OF CONVENTIONAL PCRs BASED ON PRIMERS DIRECTED TO NUCLEAR AND MITOCHONDRIAL GENES FOR THE DETECTION AND IDENTIFICATION OF Leishmania spp. |
topic_facet |
Leishmania spp kDNA Mitochondrial genes PCR Phylogeny Dogs Arctic medicine. Tropical medicine RC955-962 Infectious and parasitic diseases RC109-216 |
description |
In visceral leishmaniasis, the detection of the agent is of paramount importance to identify reservoirs of infection. Here, we evaluated the diagnostic attributes of PCRs based on primers directed to cytochrome-B (cytB), cytochrome-oxidase-subunit II (coxII), cytochrome-C (cytC), and the minicircle-kDNA. Although PCRs directed to cytB, coxII, cytC were able to detect different species of Leishmania, and the nucleotide sequence of their amplicons allowed the unequivocal differentiation of species, the analytical and diagnostic sensitivity of these PCRs were much lower than the analytical and diagnostic sensitivity of the kDNA-PCR. Among the 73 seropositive animals, the asymptomatic dogs had spleen and bone marrow samples collected and tested; only two animals were positive by PCRs based on cytB, coxII, and cytC, whereas 18 were positive by the kDNA-PCR. Considering the kDNA-PCR results, six dogs had positive spleen and bone marrow samples, eight dogs had positive bone marrow results but negative results in spleen samples and, in four dogs, the reverse situation occurred. We concluded that PCRs based on cytB, coxII, and cytC can be useful tools to identify Leishmania species when used in combination with automated sequencing. The discordance between the results of the kDNA-PCR in bone marrow and spleen samples may indicate that conventional PCR lacks sensitivity for the detection of infected dogs. Thus, primers based on the kDNA should be preferred for the screening of infected dogs. |
format |
Article in Journal/Newspaper |
author |
Estela Gallucci LOPES Carlos Alberto GERALDO JUNIOR Arlei MARCILI Ricardo Duarte SILVA Lara Borges KEID Trícia Maria Ferreira da Silva OLIVEIRA Rodrigo Martins SOARES |
author_facet |
Estela Gallucci LOPES Carlos Alberto GERALDO JUNIOR Arlei MARCILI Ricardo Duarte SILVA Lara Borges KEID Trícia Maria Ferreira da Silva OLIVEIRA Rodrigo Martins SOARES |
author_sort |
Estela Gallucci LOPES |
title |
PERFORMANCE OF CONVENTIONAL PCRs BASED ON PRIMERS DIRECTED TO NUCLEAR AND MITOCHONDRIAL GENES FOR THE DETECTION AND IDENTIFICATION OF Leishmania spp. |
title_short |
PERFORMANCE OF CONVENTIONAL PCRs BASED ON PRIMERS DIRECTED TO NUCLEAR AND MITOCHONDRIAL GENES FOR THE DETECTION AND IDENTIFICATION OF Leishmania spp. |
title_full |
PERFORMANCE OF CONVENTIONAL PCRs BASED ON PRIMERS DIRECTED TO NUCLEAR AND MITOCHONDRIAL GENES FOR THE DETECTION AND IDENTIFICATION OF Leishmania spp. |
title_fullStr |
PERFORMANCE OF CONVENTIONAL PCRs BASED ON PRIMERS DIRECTED TO NUCLEAR AND MITOCHONDRIAL GENES FOR THE DETECTION AND IDENTIFICATION OF Leishmania spp. |
title_full_unstemmed |
PERFORMANCE OF CONVENTIONAL PCRs BASED ON PRIMERS DIRECTED TO NUCLEAR AND MITOCHONDRIAL GENES FOR THE DETECTION AND IDENTIFICATION OF Leishmania spp. |
title_sort |
performance of conventional pcrs based on primers directed to nuclear and mitochondrial genes for the detection and identification of leishmania spp. |
publisher |
Universidade de São Paulo (USP) |
publishDate |
2016 |
url |
https://doi.org/10.1590/S1678-9946201658041 https://doaj.org/article/a2a8de8d60334f96971bd5c381d3105c |
geographic |
Arctic |
geographic_facet |
Arctic |
genre |
Arctic |
genre_facet |
Arctic |
op_source |
Revista do Instituto de Medicina Tropical de São Paulo, Vol 58, Iss 0 (2016) |
op_relation |
http://www.scielo.br/scielo.php?script=sci_arttext&pid=S0036-46652016005000231&lng=en&tlng=en https://doaj.org/toc/1678-9946 1678-9946 doi:10.1590/S1678-9946201658041 https://doaj.org/article/a2a8de8d60334f96971bd5c381d3105c |
op_doi |
https://doi.org/10.1590/S1678-9946201658041 |
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Revista do Instituto de Medicina Tropical de São Paulo |
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58 |
container_issue |
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