Qualitative and quantitative polymerase chain reaction (PCR) for detection of Leishmania in spleen samples from naturally infected dogs

Detalhes bibliográficos
Autor(a) principal: Solcà, Manuela da Silva
Data de Publicação: 2012
Outros Autores: Guedes, Carlos Eduardo Sampaio, Nascimento, Eliane Gomes, Oliveira, Geraldo Gileno de Sá, Santos, Washington Luis Conrado dos, Fraga, Deborah Bittencourt Mothé, Veras, Patrícia Sampaio Tavares
Tipo de documento: Artigo
Idioma: eng
Título da fonte: Repositório Institucional da UFBA
Texto Completo: http://repositorio.ufba.br/ri/handle/ri/15689
Resumo: Texto completo: acesso restrito. p. 133–140
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spelling Solcà, Manuela da SilvaGuedes, Carlos Eduardo SampaioNascimento, Eliane GomesOliveira, Geraldo Gileno de SáSantos, Washington Luis Conrado dosFraga, Deborah Bittencourt MothéVeras, Patrícia Sampaio TavaresSolcà, Manuela da SilvaGuedes, Carlos Eduardo SampaioNascimento, Eliane GomesOliveira, Geraldo Gileno de SáSantos, Washington Luis Conrado dosFraga, Deborah Bittencourt MothéVeras, Patrícia Sampaio Tavares2014-08-22T21:37:31Z20120304-4017http://repositorio.ufba.br/ri/handle/ri/15689v. 184, n. 2Texto completo: acesso restrito. p. 133–140Because infected dogs are widely considered to be the main domestic reservoir for Leishmania infantum (syn Leishmania chagasi) parasites in Brazil, the diagnosis of canine visceral leishmaniasis (CVL) must be made both accurately and promptly. The present study attempted to standardize a conventional polymerase chain reaction (cPCR) protocol for the detection of L. infantum DNA in canine spleen samples. Quantitative PCR (qPCR) technique was used to confirm the presence of Leishmania DNA in the canine spleen fragments. A comparison was made between the efficacies of these molecular diagnostic techniques and conventional parasitological and serological methods. cPCR protocols for spleen samples were standardized using primers that amplify a 145 bp fragment, located at the parasite kinetoplast minicircle. The genus specificity of the cPCR protocol was assessed by its inability to amplify the DNA of other common canine pathogens, such as Ehrlichia canis, Babesia canis, Toxoplasma gondii and Trypanosoma cruzi. cPCR protocol sensitivity was tested by assessing the reaction detection limit, determined to be 10 fg of L. infantum reference strain DNA, which corresponds to a range of 0.03–0.1 parasites per fragment. Standardized cPCR protocol was used to detect the presence of Leishmania in 45 dog spleen samples. Our results showed that 40% of the spleen fragment cultures were positive for Leishmania parasites, 58% of the dog serum samples tested positive using ELISA, and parasite DNA was detected in 44% using qPCR, while 47% of the spleen samples using cPCR. Diagnostic methods performance was assessed and revealed a better degree of ascertainment for cPCR when compared to other diagnostic methods. The sensitivity of ELISA was 83.3%, qPCR was 83.3%, and cPCR was 88.9%; PPV for ELISA was 57.7%, qPCR was 75% and cPCR was 76.2%; the Kappa coefficients were found to be 0.40 (fair) for ELISA, 0.64 (substantial) for qPCR and 0.68 (substantial) for cPCR. In both oligosymptomatic and polysymptomatic dogs, cPCR revealed the better performance analysis when compared to other diagnostic methods. The findings presented herein establish cPCR as the most indicated test to detect Leishmania when compared to the other two diagnostic methods evaluated. Despite the fact that the qPCR protocol provides a highly accurate quantification of parasites when targeting the SSU rRNA gene, this technique does not significantly improve the diagnosis of CVL when compared with the performance of the cPCR protocol, which focused on the kinetoplast minicircle.Submitted by Edileide Reis (leyde-landy@hotmail.com) on 2014-08-19T11:46:25Z No. of bitstreams: 1 Manuela da Silva Solcà.pdf: 368652 bytes, checksum: bb70de492c7851bbfe4f5d24a419bae5 (MD5)Approved for entry into archive by Patricia Barroso (pbarroso@ufba.br) on 2014-08-22T21:37:31Z (GMT) No. of bitstreams: 1 Manuela da Silva Solcà.pdf: 368652 bytes, checksum: bb70de492c7851bbfe4f5d24a419bae5 (MD5)Made available in DSpace on 2014-08-22T21:37:31Z (GMT). No. of bitstreams: 1 Manuela da Silva Solcà.pdf: 368652 bytes, checksum: bb70de492c7851bbfe4f5d24a419bae5 (MD5) Previous issue date: 2012http://dx.doi.org/ 10.1016/j.vetpar.2011.08.026reponame:Repositório Institucional da UFBAinstname:Universidade Federal da Bahia (UFBA)instacron:UFBALeishmaniaCaninePCRSpleenDiagnostic methodQualitative and quantitative polymerase chain reaction (PCR) for detection of Leishmania in spleen samples from naturally infected dogsVeterinary Parasitologyinfo:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/article10000-01-01info:eu-repo/semantics/openAccessengORIGINALManuela da Silva Solcà.pdfManuela da Silva Solcà.pdfapplication/pdf368652https://repositorio.ufba.br/bitstream/ri/15689/1/Manuela%20da%20Silva%20Solc%c3%a0.pdfbb70de492c7851bbfe4f5d24a419bae5MD51LICENSElicense.txtlicense.txttext/plain1345https://repositorio.ufba.br/bitstream/ri/15689/2/license.txtff6eaa8b858ea317fded99f125f5fcd0MD52TEXTManuela da Silva Solcà.pdf.txtManuela da Silva Solcà.pdf.txtExtracted texttext/plain46462https://repositorio.ufba.br/bitstream/ri/15689/3/Manuela%20da%20Silva%20Solc%c3%a0.pdf.txtdeaaf967c26ed74bfbe86d8dc72a7d88MD53ri/156892022-07-05 14:03:30.832oai:repositorio.ufba.br:ri/15689VGVybW8gZGUgTGljZW7vv71hLCBu77+9byBleGNsdXNpdm8sIHBhcmEgbyBkZXDvv71zaXRvIG5vIFJlcG9zaXTvv71yaW8gSW5zdGl0dWNpb25hbCBkYSBVRkJBLgoKIFBlbG8gcHJvY2Vzc28gZGUgc3VibWlzc++/vW8gZGUgZG9jdW1lbnRvcywgbyBhdXRvciBvdSBzZXUgcmVwcmVzZW50YW50ZSBsZWdhbCwgYW8gYWNlaXRhciAKZXNzZSB0ZXJtbyBkZSBsaWNlbu+/vWEsIGNvbmNlZGUgYW8gUmVwb3NpdO+/vXJpbyBJbnN0aXR1Y2lvbmFsIGRhIFVuaXZlcnNpZGFkZSBGZWRlcmFsIGRhIEJhaGlhIApvIGRpcmVpdG8gZGUgbWFudGVyIHVtYSBj77+9cGlhIGVtIHNldSByZXBvc2l077+9cmlvIGNvbSBhIGZpbmFsaWRhZGUsIHByaW1laXJhLCBkZSBwcmVzZXJ2Ye+/ve+/vW8uIApFc3NlcyB0ZXJtb3MsIG7vv71vIGV4Y2x1c2l2b3MsIG1hbnTvv71tIG9zIGRpcmVpdG9zIGRlIGF1dG9yL2NvcHlyaWdodCwgbWFzIGVudGVuZGUgbyBkb2N1bWVudG8gCmNvbW8gcGFydGUgZG8gYWNlcnZvIGludGVsZWN0dWFsIGRlc3NhIFVuaXZlcnNpZGFkZS4KCiBQYXJhIG9zIGRvY3VtZW50b3MgcHVibGljYWRvcyBjb20gcmVwYXNzZSBkZSBkaXJlaXRvcyBkZSBkaXN0cmlidWnvv73vv71vLCBlc3NlIHRlcm1vIGRlIGxpY2Vu77+9YSAKZW50ZW5kZSBxdWU6CgogTWFudGVuZG8gb3MgZGlyZWl0b3MgYXV0b3JhaXMsIHJlcGFzc2Fkb3MgYSB0ZXJjZWlyb3MsIGVtIGNhc28gZGUgcHVibGljYe+/ve+/vWVzLCBvIHJlcG9zaXTvv71yaW8KcG9kZSByZXN0cmluZ2lyIG8gYWNlc3NvIGFvIHRleHRvIGludGVncmFsLCBtYXMgbGliZXJhIGFzIGluZm9ybWHvv73vv71lcyBzb2JyZSBvIGRvY3VtZW50bwooTWV0YWRhZG9zIGVzY3JpdGl2b3MpLgoKIERlc3RhIGZvcm1hLCBhdGVuZGVuZG8gYW9zIGFuc2Vpb3MgZGVzc2EgdW5pdmVyc2lkYWRlIGVtIG1hbnRlciBzdWEgcHJvZHXvv73vv71vIGNpZW5077+9ZmljYSBjb20gCmFzIHJlc3Ryae+/ve+/vWVzIGltcG9zdGFzIHBlbG9zIGVkaXRvcmVzIGRlIHBlcmnvv71kaWNvcy4KCiBQYXJhIGFzIHB1YmxpY2Hvv73vv71lcyBzZW0gaW5pY2lhdGl2YXMgcXVlIHNlZ3VlbSBhIHBvbO+/vXRpY2EgZGUgQWNlc3NvIEFiZXJ0bywgb3MgZGVw77+9c2l0b3MgCmNvbXB1bHPvv71yaW9zIG5lc3NlIHJlcG9zaXTvv71yaW8gbWFudO+/vW0gb3MgZGlyZWl0b3MgYXV0b3JhaXMsIG1hcyBtYW5077+9bSBhY2Vzc28gaXJyZXN0cml0byAKYW8gbWV0YWRhZG9zIGUgdGV4dG8gY29tcGxldG8uIEFzc2ltLCBhIGFjZWl0Ye+/ve+/vW8gZGVzc2UgdGVybW8gbu+/vW8gbmVjZXNzaXRhIGRlIGNvbnNlbnRpbWVudG8KIHBvciBwYXJ0ZSBkZSBhdXRvcmVzL2RldGVudG9yZXMgZG9zIGRpcmVpdG9zLCBwb3IgZXN0YXJlbSBlbSBpbmljaWF0aXZhcyBkZSBhY2Vzc28gYWJlcnRvLgo=Repositório InstitucionalPUBhttp://192.188.11.11:8080/oai/requestopendoar:19322022-07-05T17:03:30Repositório Institucional da UFBA - Universidade Federal da Bahia (UFBA)false
dc.title.pt_BR.fl_str_mv Qualitative and quantitative polymerase chain reaction (PCR) for detection of Leishmania in spleen samples from naturally infected dogs
dc.title.alternative.pt_BR.fl_str_mv Veterinary Parasitology
title Qualitative and quantitative polymerase chain reaction (PCR) for detection of Leishmania in spleen samples from naturally infected dogs
spellingShingle Qualitative and quantitative polymerase chain reaction (PCR) for detection of Leishmania in spleen samples from naturally infected dogs
Solcà, Manuela da Silva
Leishmania
Canine
PCR
Spleen
Diagnostic method
title_short Qualitative and quantitative polymerase chain reaction (PCR) for detection of Leishmania in spleen samples from naturally infected dogs
title_full Qualitative and quantitative polymerase chain reaction (PCR) for detection of Leishmania in spleen samples from naturally infected dogs
title_fullStr Qualitative and quantitative polymerase chain reaction (PCR) for detection of Leishmania in spleen samples from naturally infected dogs
title_full_unstemmed Qualitative and quantitative polymerase chain reaction (PCR) for detection of Leishmania in spleen samples from naturally infected dogs
title_sort Qualitative and quantitative polymerase chain reaction (PCR) for detection of Leishmania in spleen samples from naturally infected dogs
author Solcà, Manuela da Silva
author_facet Solcà, Manuela da Silva
Guedes, Carlos Eduardo Sampaio
Nascimento, Eliane Gomes
Oliveira, Geraldo Gileno de Sá
Santos, Washington Luis Conrado dos
Fraga, Deborah Bittencourt Mothé
Veras, Patrícia Sampaio Tavares
author_role author
author2 Guedes, Carlos Eduardo Sampaio
Nascimento, Eliane Gomes
Oliveira, Geraldo Gileno de Sá
Santos, Washington Luis Conrado dos
Fraga, Deborah Bittencourt Mothé
Veras, Patrícia Sampaio Tavares
author2_role author
author
author
author
author
author
dc.contributor.author.fl_str_mv Solcà, Manuela da Silva
Guedes, Carlos Eduardo Sampaio
Nascimento, Eliane Gomes
Oliveira, Geraldo Gileno de Sá
Santos, Washington Luis Conrado dos
Fraga, Deborah Bittencourt Mothé
Veras, Patrícia Sampaio Tavares
Solcà, Manuela da Silva
Guedes, Carlos Eduardo Sampaio
Nascimento, Eliane Gomes
Oliveira, Geraldo Gileno de Sá
Santos, Washington Luis Conrado dos
Fraga, Deborah Bittencourt Mothé
Veras, Patrícia Sampaio Tavares
dc.subject.por.fl_str_mv Leishmania
Canine
PCR
Spleen
Diagnostic method
topic Leishmania
Canine
PCR
Spleen
Diagnostic method
description Texto completo: acesso restrito. p. 133–140
publishDate 2012
dc.date.issued.fl_str_mv 2012
dc.date.accessioned.fl_str_mv 2014-08-22T21:37:31Z
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dc.identifier.uri.fl_str_mv http://repositorio.ufba.br/ri/handle/ri/15689
dc.identifier.issn.none.fl_str_mv 0304-4017
dc.identifier.number.pt_BR.fl_str_mv v. 184, n. 2
identifier_str_mv 0304-4017
v. 184, n. 2
url http://repositorio.ufba.br/ri/handle/ri/15689
dc.language.iso.fl_str_mv eng
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