Comparison of phenol-chloroform and a commercial deoxyribonucleic acid extraction kit for identification of bloodmeal sources from triatomines (Hemiptera: Reduviidae)

Detalhes bibliográficos
Autor(a) principal: Silva, Andressa Noronha Barbosa da
Data de Publicação: 2020
Outros Autores: Souza, Rita de Cássia Moreira de, Honorato, Nathan Ravi Medeiros, Martins, Rand Randall, Câmara, Antônia Claudia Jácome da, Galvão, Lúcia Maria da Cunha, Chiari, Egler
Tipo de documento: Artigo
Idioma: eng
Título da fonte: Repositório Institucional da FIOCRUZ (ARCA)
Texto Completo: https://www.arca.fiocruz.br/handle/icict/58732
Resumo: Universidade Federal do Rio Grande do Norte. Centro de Ciências da Saúde. Programa de Pós-Graduação em Ciências Farmacêuticas. Natal, RN, Brasil/Universidade Federal de Minas Gerais. Instituto de Ciências Biológicas. Programa de Pós-Graduação em Parasitologia. Belo Horizonte, MG, Brasil.
id CRUZ_99d6754172fef3c6432b51c9f57d37db
oai_identifier_str oai:www.arca.fiocruz.br:icict/58732
network_acronym_str CRUZ
network_name_str Repositório Institucional da FIOCRUZ (ARCA)
repository_id_str 2135
spelling Silva, Andressa Noronha Barbosa daSouza, Rita de Cássia Moreira deHonorato, Nathan Ravi MedeirosMartins, Rand RandallCâmara, Antônia Claudia Jácome daGalvão, Lúcia Maria da CunhaChiari, Egler2023-05-25T19:48:57Z2023-05-25T19:48:57Z2020SILVA, Andressa Noronha Barbosa da et al. Comparison of phenol-chloroform and a commercial deoxyribonucleic acid extraction kit for identification of bloodmeal sources from triatomines (Hemiptera: Reduviidae). Rev Soc Bras Med Trop., v. 53, e20200189, 2020. doi: 10.1590/0037-8682-0189-2020.0037-8682https://www.arca.fiocruz.br/handle/icict/5873210.1590/0037-8682-0189-2020engSociedade Brasileira de Medicina TropicalComparison of phenol-chloroform and a commercial deoxyribonucleic acid extraction kit for identification of bloodmeal sources from triatomines (Hemiptera: Reduviidae)info:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/articleUniversidade Federal do Rio Grande do Norte. Centro de Ciências da Saúde. Programa de Pós-Graduação em Ciências Farmacêuticas. Natal, RN, Brasil/Universidade Federal de Minas Gerais. Instituto de Ciências Biológicas. Programa de Pós-Graduação em Parasitologia. Belo Horizonte, MG, Brasil.Fundação Oswaldo Cruz. Instituto René Rachou. Belo Horizonte, MG, Brasil.Universidade Federal do Rio Grande do Norte. Centro de Biociências. Programa de Pós-Graduação em Biologia Parasitária. Natal, RN, Brasil.Universidade Federal do Rio Grande do Norte. Centro de Ciências da Saúde. Programa de Pós-Graduação em Ciências Aplicadas à Saúde da Mulher. Natal, RN, Brasil.Universidade Federal do Rio Grande do Norte. Centro de Ciências da Saúde. Programa de Pós-Graduação em Ciências Farmacêuticas.Natal, RN, Brasil/Universidade Federal do Rio Grande do Norte. Centro de Biociências. Programa de Pós-Graduação em Biologia Parasitária. Natal, RN, Brasil.Universidade Federal do Rio Grande do Norte. Centro de Ciências da Saúde. Programa de Pós-Graduação em Ciências Farmacêuticas. Natal, RN, Brasil/Universidade Federal de Minas Gerais. Instituto de Ciências Biológicas. Programa de Pós-Graduação em Parasitologia. Belo Horizonte, MG, Brasil.Universidade Federal de Minas Gerais. Instituto de Ciências Biológicas. Programa de Pós-Graduação em Parasitologia. Belo Horizonte, MG, Brasil.Introduction: Knowledge of triatomine bloodmeal sources is essential for understanding vector-host interactions in Trypanosoma cruzi transmission cycles. Expensive commercial deoxyribonucleic acid (DNA) extraction kits are widely used for bloodmeal identification. This study assessed the performance of an inexpensive phenol-chloroform DNA extraction protocol for identification of triatomine bloodmeal sources, comparing it with a conunercially available kit. Methods: Both methods were used to obtain DNA from the intestinal contents of Triatoma brasiliensis blood-fed on either Columba sp., Alus musculus, or Gallus gallus. Subsequently, the mitochondria] 12S ribosomal ribonucleic acid (rRNA) gene was amplified by polymerase chain reaction, sequenced, and compared with GenBank data. Results: Twelve (80%) samples extracted with the commercial kit and four (26.7%) with phenol-chloroform were pure (according to the A260/A280 ratio). Samples extracted with phenol-chloroform, except for Columba sp. samples, had higher DNA concentration than those extracted with the commercial kit. Samples extracted using phenol-chloroform and blood-fed on G. gallus had significantly higher DNA concentration than those blood-fed on Columba sp. (p-value <0.001) and M musculus (p-value <0.001). The 215-base-pair 12S rRNA fragment was amplified from all samples and produced reliable sequences, enabling the identification of the bloodmeal source, most of which showed identity and coverage above 95%. The phenol-chloroform method was much less expensive than the commercial kit but took considerably more time to perform. Conclusions: Our data showed that both DNA extraction methods produced reliable sequences enabling identification of triatomine bloodmeal sources but differed greatly in cost and time required.TriatominaeGastrointestinal contentsRNA ribosomal 12SPolymerase Chain Reactioninfo:eu-repo/semantics/openAccessreponame:Repositório Institucional da FIOCRUZ (ARCA)instname:Fundação Oswaldo Cruz (FIOCRUZ)instacron:FIOCRUZORIGINALA_0189_2020.indd.pdfA_0189_2020.indd.pdfapplication/pdf1320185https://www.arca.fiocruz.br/bitstream/icict/58732/2/A_0189_2020.indd.pdfef6562eb2d589edb4de2f4764c42b879MD52LICENSElicense.txttext/plain1748https://www.arca.fiocruz.br/bitstream/icict/58732/1/license.txt8a4605be74aa9ea9d79846c1fba20a33MD51icict/587322023-05-25 16:48:57.979oai:www.arca.fiocruz.br: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Repositório InstitucionalPUBhttps://www.arca.fiocruz.br/oai/requestrepositorio.arca@fiocruz.bropendoar:21352023-05-25T19:48:57Repositório Institucional da FIOCRUZ (ARCA) - Fundação Oswaldo Cruz (FIOCRUZ)false
dc.title.en_US.fl_str_mv Comparison of phenol-chloroform and a commercial deoxyribonucleic acid extraction kit for identification of bloodmeal sources from triatomines (Hemiptera: Reduviidae)
title Comparison of phenol-chloroform and a commercial deoxyribonucleic acid extraction kit for identification of bloodmeal sources from triatomines (Hemiptera: Reduviidae)
spellingShingle Comparison of phenol-chloroform and a commercial deoxyribonucleic acid extraction kit for identification of bloodmeal sources from triatomines (Hemiptera: Reduviidae)
Silva, Andressa Noronha Barbosa da
Triatominae
Gastrointestinal contents
RNA ribosomal 12S
Polymerase Chain Reaction
title_short Comparison of phenol-chloroform and a commercial deoxyribonucleic acid extraction kit for identification of bloodmeal sources from triatomines (Hemiptera: Reduviidae)
title_full Comparison of phenol-chloroform and a commercial deoxyribonucleic acid extraction kit for identification of bloodmeal sources from triatomines (Hemiptera: Reduviidae)
title_fullStr Comparison of phenol-chloroform and a commercial deoxyribonucleic acid extraction kit for identification of bloodmeal sources from triatomines (Hemiptera: Reduviidae)
title_full_unstemmed Comparison of phenol-chloroform and a commercial deoxyribonucleic acid extraction kit for identification of bloodmeal sources from triatomines (Hemiptera: Reduviidae)
title_sort Comparison of phenol-chloroform and a commercial deoxyribonucleic acid extraction kit for identification of bloodmeal sources from triatomines (Hemiptera: Reduviidae)
author Silva, Andressa Noronha Barbosa da
author_facet Silva, Andressa Noronha Barbosa da
Souza, Rita de Cássia Moreira de
Honorato, Nathan Ravi Medeiros
Martins, Rand Randall
Câmara, Antônia Claudia Jácome da
Galvão, Lúcia Maria da Cunha
Chiari, Egler
author_role author
author2 Souza, Rita de Cássia Moreira de
Honorato, Nathan Ravi Medeiros
Martins, Rand Randall
Câmara, Antônia Claudia Jácome da
Galvão, Lúcia Maria da Cunha
Chiari, Egler
author2_role author
author
author
author
author
author
dc.contributor.author.fl_str_mv Silva, Andressa Noronha Barbosa da
Souza, Rita de Cássia Moreira de
Honorato, Nathan Ravi Medeiros
Martins, Rand Randall
Câmara, Antônia Claudia Jácome da
Galvão, Lúcia Maria da Cunha
Chiari, Egler
dc.subject.en.en_US.fl_str_mv Triatominae
Gastrointestinal contents
RNA ribosomal 12S
Polymerase Chain Reaction
topic Triatominae
Gastrointestinal contents
RNA ribosomal 12S
Polymerase Chain Reaction
description Universidade Federal do Rio Grande do Norte. Centro de Ciências da Saúde. Programa de Pós-Graduação em Ciências Farmacêuticas. Natal, RN, Brasil/Universidade Federal de Minas Gerais. Instituto de Ciências Biológicas. Programa de Pós-Graduação em Parasitologia. Belo Horizonte, MG, Brasil.
publishDate 2020
dc.date.issued.fl_str_mv 2020
dc.date.accessioned.fl_str_mv 2023-05-25T19:48:57Z
dc.date.available.fl_str_mv 2023-05-25T19:48:57Z
dc.type.status.fl_str_mv info:eu-repo/semantics/publishedVersion
dc.type.driver.fl_str_mv info:eu-repo/semantics/article
format article
status_str publishedVersion
dc.identifier.citation.fl_str_mv SILVA, Andressa Noronha Barbosa da et al. Comparison of phenol-chloroform and a commercial deoxyribonucleic acid extraction kit for identification of bloodmeal sources from triatomines (Hemiptera: Reduviidae). Rev Soc Bras Med Trop., v. 53, e20200189, 2020. doi: 10.1590/0037-8682-0189-2020.
dc.identifier.uri.fl_str_mv https://www.arca.fiocruz.br/handle/icict/58732
dc.identifier.issn.en_US.fl_str_mv 0037-8682
dc.identifier.doi.none.fl_str_mv 10.1590/0037-8682-0189-2020
identifier_str_mv SILVA, Andressa Noronha Barbosa da et al. Comparison of phenol-chloroform and a commercial deoxyribonucleic acid extraction kit for identification of bloodmeal sources from triatomines (Hemiptera: Reduviidae). Rev Soc Bras Med Trop., v. 53, e20200189, 2020. doi: 10.1590/0037-8682-0189-2020.
0037-8682
10.1590/0037-8682-0189-2020
url https://www.arca.fiocruz.br/handle/icict/58732
dc.language.iso.fl_str_mv eng
language eng
dc.rights.driver.fl_str_mv info:eu-repo/semantics/openAccess
eu_rights_str_mv openAccess
dc.publisher.none.fl_str_mv Sociedade Brasileira de Medicina Tropical
publisher.none.fl_str_mv Sociedade Brasileira de Medicina Tropical
dc.source.none.fl_str_mv reponame:Repositório Institucional da FIOCRUZ (ARCA)
instname:Fundação Oswaldo Cruz (FIOCRUZ)
instacron:FIOCRUZ
instname_str Fundação Oswaldo Cruz (FIOCRUZ)
instacron_str FIOCRUZ
institution FIOCRUZ
reponame_str Repositório Institucional da FIOCRUZ (ARCA)
collection Repositório Institucional da FIOCRUZ (ARCA)
bitstream.url.fl_str_mv https://www.arca.fiocruz.br/bitstream/icict/58732/2/A_0189_2020.indd.pdf
https://www.arca.fiocruz.br/bitstream/icict/58732/1/license.txt
bitstream.checksum.fl_str_mv ef6562eb2d589edb4de2f4764c42b879
8a4605be74aa9ea9d79846c1fba20a33
bitstream.checksumAlgorithm.fl_str_mv MD5
MD5
repository.name.fl_str_mv Repositório Institucional da FIOCRUZ (ARCA) - Fundação Oswaldo Cruz (FIOCRUZ)
repository.mail.fl_str_mv repositorio.arca@fiocruz.br
_version_ 1813008838085836800

Registros relacionados