Accuracy of quantitative polymerase chain reaction in samples of frozen and paraffin-embedded healthy skin for the diagnosis of canine visceral leishmaniasis

ABSTRACT The purpose of the present work was to evaluate the accuracy of quantitative polymerase chain reaction (qPCR) performed on samples of fresh frozen tissue (FT) and formalin-fixed, paraffin-embedded (FFPE) healthy skin. This is a validation study conducted with samples from 46 dogs from an en...

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Main Authors: M.P. Campos, M.F. Madeira, D.A. Silva, M.S. Solcà, O.M. Espíndola, A.A.V. Mendes Júnior, F.B. Figueiredo
Format: Article
Language:English
Published: Universidade Federal de Minas Gerais
Series:Arquivo Brasileiro de Medicina Veterinária e Zootecnia
Subjects:
Online Access:http://www.scielo.br/scielo.php?script=sci_arttext&pid=S0102-09352017000601443&lng=en&tlng=en
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author M.P. Campos
M.F. Madeira
D.A. Silva
M.S. Solcà
O.M. Espíndola
A.A.V. Mendes Júnior
F.B. Figueiredo
author_facet M.P. Campos
M.F. Madeira
D.A. Silva
M.S. Solcà
O.M. Espíndola
A.A.V. Mendes Júnior
F.B. Figueiredo
author_sort M.P. Campos
collection DOAJ
description ABSTRACT The purpose of the present work was to evaluate the accuracy of quantitative polymerase chain reaction (qPCR) performed on samples of fresh frozen tissue (FT) and formalin-fixed, paraffin-embedded (FFPE) healthy skin. This is a validation study conducted with samples from 46 dogs from an endemic area in Brazil. After sample collection, DNA extractions were conducted using commercial kits and qPCR was oriented to kinetoplast DNA (kDNA) targets of the Leishmania infantum species. The results obtained for the FFPE samples showed 63.6% sensitivity and 77.1% specificity, whereas those obtained for the FT samples showed 100% and 48.6%, respectively. Poor agreement was observed for the results of the qPCR technique with FT and FFPE samples. Our results suggest freezing as the most suitable conservation method for the formation of sample databases considering DNA recovery
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spelling doaj.art-714b481d873e4bda971637ccc42876b42022-12-22T00:09:03ZengUniversidade Federal de Minas GeraisArquivo Brasileiro de Medicina Veterinária e Zootecnia1678-41626961443145010.1590/1678-4162-9053S0102-09352017000601443Accuracy of quantitative polymerase chain reaction in samples of frozen and paraffin-embedded healthy skin for the diagnosis of canine visceral leishmaniasisM.P. CamposM.F. MadeiraD.A. SilvaM.S. SolcàO.M. EspíndolaA.A.V. Mendes JúniorF.B. FigueiredoABSTRACT The purpose of the present work was to evaluate the accuracy of quantitative polymerase chain reaction (qPCR) performed on samples of fresh frozen tissue (FT) and formalin-fixed, paraffin-embedded (FFPE) healthy skin. This is a validation study conducted with samples from 46 dogs from an endemic area in Brazil. After sample collection, DNA extractions were conducted using commercial kits and qPCR was oriented to kinetoplast DNA (kDNA) targets of the Leishmania infantum species. The results obtained for the FFPE samples showed 63.6% sensitivity and 77.1% specificity, whereas those obtained for the FT samples showed 100% and 48.6%, respectively. Poor agreement was observed for the results of the qPCR technique with FT and FFPE samples. Our results suggest freezing as the most suitable conservation method for the formation of sample databases considering DNA recoveryhttp://www.scielo.br/scielo.php?script=sci_arttext&pid=S0102-09352017000601443&lng=en&tlng=endiagnósticoqPCRleishmaniose visceral canina
spellingShingle M.P. Campos
M.F. Madeira
D.A. Silva
M.S. Solcà
O.M. Espíndola
A.A.V. Mendes Júnior
F.B. Figueiredo
Accuracy of quantitative polymerase chain reaction in samples of frozen and paraffin-embedded healthy skin for the diagnosis of canine visceral leishmaniasis
Arquivo Brasileiro de Medicina Veterinária e Zootecnia
diagnóstico
qPCR
leishmaniose visceral canina
title Accuracy of quantitative polymerase chain reaction in samples of frozen and paraffin-embedded healthy skin for the diagnosis of canine visceral leishmaniasis
title_full Accuracy of quantitative polymerase chain reaction in samples of frozen and paraffin-embedded healthy skin for the diagnosis of canine visceral leishmaniasis
title_fullStr Accuracy of quantitative polymerase chain reaction in samples of frozen and paraffin-embedded healthy skin for the diagnosis of canine visceral leishmaniasis
title_full_unstemmed Accuracy of quantitative polymerase chain reaction in samples of frozen and paraffin-embedded healthy skin for the diagnosis of canine visceral leishmaniasis
title_short Accuracy of quantitative polymerase chain reaction in samples of frozen and paraffin-embedded healthy skin for the diagnosis of canine visceral leishmaniasis
title_sort accuracy of quantitative polymerase chain reaction in samples of frozen and paraffin embedded healthy skin for the diagnosis of canine visceral leishmaniasis
topic diagnóstico
qPCR
leishmaniose visceral canina
url http://www.scielo.br/scielo.php?script=sci_arttext&pid=S0102-09352017000601443&lng=en&tlng=en
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