Qualitative and quantitative polymerase chain reaction (PCR) for detection of Leishmania in spleen samples from naturally infected dogs
| Autor(a) principal: | |
|---|---|
| Data de Publicação: | 2012 |
| Outros Autores: | , , , , , |
| Tipo de documento: | Artigo |
| Idioma: | eng |
| Título da fonte: | Repositório Institucional da FIOCRUZ (ARCA) |
| Texto Completo: | https://www.arca.fiocruz.br/handle/icict/7305 |
Resumo: | Fundação Oswaldo Cruz. Centro de Pesquisa Gonçalo Moniz. Laboratório de Patologia e Biointervenção. Salvador, BA, Brasil |
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Solcà, Manuela da SilvaGuedes, Carlos Eduardo SampaioNascimento, Eliane GomesOliveira, Geraldo Gileno de SádosSantos, Washington Luis ConradoFraga, Deborah Bittencourt MothéVeras, Patrícia Sampaio Tavares2014-02-10T13:16:03Z2014-02-10T13:16:03Z2012SOLCA, M. da S. et al. Qualitative and quantitative polymerase chain reaction (PCR) for detection of Leishmania in spleen samples from naturally infected dogs. Veterinary Parasitology, v. 23, n. 2-4, p. 133-140, 2012.0304-4017https://www.arca.fiocruz.br/handle/icict/730510.1016/j.vetpar.2011.08.026engElsevierQualitative and quantitative polymerase chain reaction (PCR) for detection of Leishmania in spleen samples from naturally infected dogsinfo:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/articleFundação Oswaldo Cruz. Centro de Pesquisa Gonçalo Moniz. Laboratório de Patologia e Biointervenção. Salvador, BA, BrasilFundação Oswaldo Cruz. Centro de Pesquisa Gonçalo Moniz. Laboratório de Patologia e Biointervenção. Salvador, BA, BrasilCentro de Referência em Doenças Endêmicas Pirajá da Silva-PIEJ. Jequié, Ba, BrasilFundação Oswaldo Cruz. Centro de Pesquisa Gonçalo Moniz. Laboratório de Patologia e Biointervenção. Salvador, BA, BrasilFundação Oswaldo Cruz. Centro de Pesquisa Gonçalo Moniz. Laboratório de Patologia e Biointervenção. Salvador, BA, BrasilFundação Oswaldo Cruz. Centro de Pesquisa Gonçalo Moniz. Laboratório de Patologia e Biointervenção. Salvador, BA, Brasil / Universidade Federal da Bahia. Escola de Medicina Veterinária. Departamento de Medicina Veterinária Preventiva. Salvador, BA, BrasilFundação Oswaldo Cruz. Centro de Pesquisa Gonçalo Moniz. Laboratório de Patologia e Biointervenção. Salvador, BA, BrasilBecause 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 minicircleLeishmaniaCaninePCRSpleenDiagnostic methodDoenças do Cão/diagnósticoLeishmania/genéticaLeishmaniose/veterináriaReação em Cadeia da Polimerase/veterináriaBaço/parasitologiaAnimaisPrimers do DNA/genéticaDoenças do Cão/parasitologiaCãesLeishmaniose/diagnósticoLeishmaniose/parasitologiaSensibilidade e Especificidadeinfo:eu-repo/semantics/openAccessreponame:Repositório Institucional da FIOCRUZ (ARCA)instname:Fundação Oswaldo Cruz (FIOCRUZ)instacron:FIOCRUZORIGINALSolcà, Mda S. et al. Qualitative and quantitative. Vet Parasitol 2012; v. 184,p. 185.pdfSolcà, Mda S. et al. Qualitative and quantitative. Vet Parasitol 2012; v. 184,p. 185.pdfapplication/pdf368652https://www.arca.fiocruz.br/bitstream/icict/7305/1/Solc%c3%a0%2c%20Mda%20S.%20et%20al.%20Qualitative%20and%20quantitative.%20Vet%20Parasitol%202012%3b%20v.%20184%2cp.%20185.pdfbb70de492c7851bbfe4f5d24a419bae5MD51LICENSElicense.txtlicense.txttext/plain; charset=utf-81914https://www.arca.fiocruz.br/bitstream/icict/7305/2/license.txt7d48279ffeed55da8dfe2f8e81f3b81fMD52TEXTSolcà, Mda S. et al. Qualitative and quantitative. Vet Parasitol 2012; v. 184,p. 185.pdf.txtSolcà, Mda S. et al. Qualitative and quantitative. Vet Parasitol 2012; v. 184,p. 185.pdf.txtExtracted texttext/plain47218https://www.arca.fiocruz.br/bitstream/icict/7305/5/Solc%c3%a0%2c%20Mda%20S.%20et%20al.%20Qualitative%20and%20quantitative.%20Vet%20Parasitol%202012%3b%20v.%20184%2cp.%20185.pdf.txt5f0b608d8ee27b94b613f97d879978b0MD55THUMBNAILSolcà, Mda S. et al. 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| 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 |
| 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 Doenças do Cão/diagnóstico Leishmania/genética Leishmaniose/veterinária Reação em Cadeia da Polimerase/veterinária Baço/parasitologia Animais Primers do DNA/genética Doenças do Cão/parasitologia Cães Leishmaniose/diagnóstico Leishmaniose/parasitologia Sensibilidade e Especificidade |
| 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 dosSantos, Washington Luis Conrado Fraga, Deborah Bittencourt Mothé Veras, Patrícia Sampaio Tavares |
| author_role |
author |
| author2 |
Guedes, Carlos Eduardo Sampaio Nascimento, Eliane Gomes Oliveira, Geraldo Gileno de Sá dosSantos, Washington Luis Conrado 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 dosSantos, Washington Luis Conrado Fraga, Deborah Bittencourt Mothé Veras, Patrícia Sampaio Tavares |
| dc.subject.en.pt_BR.fl_str_mv |
Leishmania Canine PCR Spleen Diagnostic method |
| topic |
Leishmania Canine PCR Spleen Diagnostic method Doenças do Cão/diagnóstico Leishmania/genética Leishmaniose/veterinária Reação em Cadeia da Polimerase/veterinária Baço/parasitologia Animais Primers do DNA/genética Doenças do Cão/parasitologia Cães Leishmaniose/diagnóstico Leishmaniose/parasitologia Sensibilidade e Especificidade |
| dc.subject.decs.pt_BR.fl_str_mv |
Doenças do Cão/diagnóstico Leishmania/genética Leishmaniose/veterinária Reação em Cadeia da Polimerase/veterinária Baço/parasitologia Animais Primers do DNA/genética Doenças do Cão/parasitologia Cães Leishmaniose/diagnóstico Leishmaniose/parasitologia Sensibilidade e Especificidade |
| description |
Fundação Oswaldo Cruz. Centro de Pesquisa Gonçalo Moniz. Laboratório de Patologia e Biointervenção. Salvador, BA, Brasil |
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2012 |
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2012 |
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2014-02-10T13:16:03Z |
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2014-02-10T13:16:03Z |
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info:eu-repo/semantics/publishedVersion |
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info:eu-repo/semantics/article |
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publishedVersion |
| dc.identifier.citation.fl_str_mv |
SOLCA, M. da S. et al. Qualitative and quantitative polymerase chain reaction (PCR) for detection of Leishmania in spleen samples from naturally infected dogs. Veterinary Parasitology, v. 23, n. 2-4, p. 133-140, 2012. |
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https://www.arca.fiocruz.br/handle/icict/7305 |
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0304-4017 |
| dc.identifier.doi.none.fl_str_mv |
10.1016/j.vetpar.2011.08.026 |
| identifier_str_mv |
SOLCA, M. da S. et al. Qualitative and quantitative polymerase chain reaction (PCR) for detection of Leishmania in spleen samples from naturally infected dogs. Veterinary Parasitology, v. 23, n. 2-4, p. 133-140, 2012. 0304-4017 10.1016/j.vetpar.2011.08.026 |
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Elsevier |
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Elsevier |
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