PCR-RFLP screening of polymorphisms associated with benzimidazole resistance in Necator americanus and Ascaris lumbricoides from different geographical regions in Brazil

Detalhes bibliográficos
Autor(a) principal: Luciana Werneck Zuccherato
Data de Publicação: 2018
Outros Autores: Luiz Fernando Vieira Furtado, Celi da Silva Medeiros, Carina da Silva Pinheiro, Elida Mara Leite Rabelo
Tipo de documento: Artigo
Idioma: eng
Título da fonte: Repositório Institucional da UFMG
Texto Completo: https://doi.org/10.1371/journal.pntd.0006766
http://hdl.handle.net/1843/39486
Resumo: Ascaris lumbricoides and Necator americanus are soil-transmitted parasites with global geographic distribution, and they represent some of the most common and neglected infec tions in the world. Periodic treatment with mass drug administration (MDA) in endemic areas is the recommended action put forth by the World Health Organization. However, MDA can cause the selection of subpopulations that possess the genetic ability to overcome the mechanism of drug action. In fact, beta-tubulin gene mutations (codons 167, 198 and 200) are correlated with benzimidazole resistance in nematodes of veterinary importance. It is possible that these SNPs also have strong correlation with treatment resistance in the human geohelminths A. lumbricoides, Trichuris trichiura and hookworms. Here, we aimed to investigate the presence of some of these canonical molecular markers associated with par asite resistance to benzimidazole in N. americanus and A. lumbricoides collected from six Brazilian states. Nested-PCR and PCR-RFLP were used to detect mutations at codons 167 and 198 in 601 individual eggs of A. lumbricoides collected from 62 human stool samples; however, no mutations were found. Codons 198 and 200 were tested in 552 N. americanus eggs collected from 48 patients using the same methodology, which presented a relative fre quency of 1.4% and 1.1%, respectively. The presence of these SNPs in N. americanus eggs is an important finding, indicating that with high benzimidazole drug pressure there is potential for benzimidazole resistance to be selected in this hookworm. However, at these low frequencies it does not indicate that there is at present any benzimidazole resistance problem. This is the first systematic study performed in South America, and the study yielded a landscape of the genetic variants in the beta-tubulin gene and anthelmintic resis tance to soil-transmitted parasites detected by a simple, rapid and affordable genotyping assay of individual eggs
id UFMG_ab071032bbcb5610f291912be05e557a
oai_identifier_str oai:repositorio.ufmg.br:1843/39486
network_acronym_str UFMG
network_name_str Repositório Institucional da UFMG
repository_id_str
spelling 2022-02-18T21:47:14Z2022-02-18T21:47:14Z2018-09-1712113https://doi.org/10.1371/journal.pntd.000676603786978http://hdl.handle.net/1843/39486Ascaris lumbricoides and Necator americanus are soil-transmitted parasites with global geographic distribution, and they represent some of the most common and neglected infec tions in the world. Periodic treatment with mass drug administration (MDA) in endemic areas is the recommended action put forth by the World Health Organization. However, MDA can cause the selection of subpopulations that possess the genetic ability to overcome the mechanism of drug action. In fact, beta-tubulin gene mutations (codons 167, 198 and 200) are correlated with benzimidazole resistance in nematodes of veterinary importance. It is possible that these SNPs also have strong correlation with treatment resistance in the human geohelminths A. lumbricoides, Trichuris trichiura and hookworms. Here, we aimed to investigate the presence of some of these canonical molecular markers associated with par asite resistance to benzimidazole in N. americanus and A. lumbricoides collected from six Brazilian states. Nested-PCR and PCR-RFLP were used to detect mutations at codons 167 and 198 in 601 individual eggs of A. lumbricoides collected from 62 human stool samples; however, no mutations were found. Codons 198 and 200 were tested in 552 N. americanus eggs collected from 48 patients using the same methodology, which presented a relative fre quency of 1.4% and 1.1%, respectively. The presence of these SNPs in N. americanus eggs is an important finding, indicating that with high benzimidazole drug pressure there is potential for benzimidazole resistance to be selected in this hookworm. However, at these low frequencies it does not indicate that there is at present any benzimidazole resistance problem. This is the first systematic study performed in South America, and the study yielded a landscape of the genetic variants in the beta-tubulin gene and anthelmintic resis tance to soil-transmitted parasites detected by a simple, rapid and affordable genotyping assay of individual eggsAscaris lumbricoides e Necator americanus são parasitas transmitidos pelo solo com distribuição geográfica, e representam algumas das infecções mais comuns e negligenciadas no mundo. Tratamento periódico com administração em massa de medicamentos (MDA) em áreas endêmicas é a ação recomendada pela Organização Mundial da Saúde. No entanto, o MDA pode causar a seleção de subpopulações que possuem a capacidade genética de superar o mecanismo de ação da droga. De fato, mutações no gene da beta-tubulina (códons 167, 198 e 200) estão correlacionados com a resistência ao benzimidazol em nematoides de importância veterinária. Isto é possível que esses SNPs também tenham forte correlação com a resistência ao tratamento no geohelmintos humanos A. lumbricoides, Trichuris trichiura e ancilostomídeos. Aqui, tivemos como objetivo investigar a presença de alguns desses marcadores moleculares canônicos associados à resistência parasitária ao benzimidazol em N. americanus e A. lumbricoides coletados de seis estados brasileiros. Nested-PCR e PCR-RFLP foram usados ​​para detectar mutações nos códons 167 e 198 em 601 ovos individuais de A. lumbricoides coletados de 62 amostras de fezes humanas; no entanto, nenhuma mutação foi encontrada. Os códons 198 e 200 foram testados em 552 N. americanus ovos coletados de 48 pacientes pela mesma metodologia, que apresentaram frequência relativa de 1,4% e 1,1%, respectivamente. A presença desses SNPs em N. americanus ovos é um achado importante, indicando que com alta pressão do medicamento benzimidazol há potencial de resistência ao benzimidazol a ser selecionado neste ancilostomídeo. No entanto, nestas baixas frequências, isso não indica que exista atualmente qualquer resistência ao benzimidazol problema. Este é o primeiro estudo sistemático realizado na América do Sul, e o estudo produziu um panorama das variantes genéticas no gene da beta-tubulina e resistência anti-helmíntica a parasitas transmitidos pelo solo detectados por uma genotipagem simples, rápida e acessível ensaio de ovos individuaisengUniversidade Federal de Minas GeraisUFMGBrasilICB - INSTITUTO DE CIÊNCIAS BIOLOGICASPlos neglected tropical diseasesAscaris lumbricoidesNecator americanusPCR-RFLP screening of polymorphisms associated with benzimidazole resistance in Necator americanus and Ascaris lumbricoides from different geographical regions in BrazilTriagem PCR-RFLP de polimorfismos associada à resistência ao benzimidazol em Necator americanus e Ascaris lumbricoides de diferentes regiões geográficas do Brasilinfo:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/articleLuciana Werneck ZuccheratoLuiz Fernando Vieira FurtadoCeli da Silva MedeirosCarina da Silva PinheiroElida Mara Leite Rabeloapplication/pdfinfo:eu-repo/semantics/openAccessreponame:Repositório Institucional da UFMGinstname:Universidade Federal de Minas Gerais (UFMG)instacron:UFMGLICENSELicense.txtLicense.txttext/plain; charset=utf-82042https://repositorio.ufmg.br/bitstream/1843/39486/1/License.txtfa505098d172de0bc8864fc1287ffe22MD51ORIGINALPCR-RFLP_screening_of_polymorphisms_associated_wit.pdfPCR-RFLP_screening_of_polymorphisms_associated_wit.pdfapplication/pdf1685495https://repositorio.ufmg.br/bitstream/1843/39486/2/PCR-RFLP_screening_of_polymorphisms_associated_wit.pdf734f26661c94c65e0760b0cc0f2bd6faMD521843/394862022-02-18 18:47:14.687oai:repositorio.ufmg.br:1843/39486TElDRU7vv71BIERFIERJU1RSSUJVSe+/ve+/vU8gTu+/vU8tRVhDTFVTSVZBIERPIFJFUE9TSVTvv71SSU8gSU5TVElUVUNJT05BTCBEQSBVRk1HCiAKCkNvbSBhIGFwcmVzZW50Ye+/ve+/vW8gZGVzdGEgbGljZW7vv71hLCB2b2Pvv70gKG8gYXV0b3IgKGVzKSBvdSBvIHRpdHVsYXIgZG9zIGRpcmVpdG9zIGRlIGF1dG9yKSBjb25jZWRlIGFvIFJlcG9zaXTvv71yaW8gSW5zdGl0dWNpb25hbCBkYSBVRk1HIChSSS1VRk1HKSBvIGRpcmVpdG8gbu+/vW8gZXhjbHVzaXZvIGUgaXJyZXZvZ++/vXZlbCBkZSByZXByb2R1emlyIGUvb3UgZGlzdHJpYnVpciBhIHN1YSBwdWJsaWNh77+977+9byAoaW5jbHVpbmRvIG8gcmVzdW1vKSBwb3IgdG9kbyBvIG11bmRvIG5vIGZvcm1hdG8gaW1wcmVzc28gZSBlbGV0cu+/vW5pY28gZSBlbSBxdWFscXVlciBtZWlvLCBpbmNsdWluZG8gb3MgZm9ybWF0b3Mg77+9dWRpbyBvdSB277+9ZGVvLgoKVm9j77+9IGRlY2xhcmEgcXVlIGNvbmhlY2UgYSBwb2zvv710aWNhIGRlIGNvcHlyaWdodCBkYSBlZGl0b3JhIGRvIHNldSBkb2N1bWVudG8gZSBxdWUgY29uaGVjZSBlIGFjZWl0YSBhcyBEaXJldHJpemVzIGRvIFJJLVVGTUcuCgpWb2Pvv70gY29uY29yZGEgcXVlIG8gUmVwb3NpdO+/vXJpbyBJbnN0aXR1Y2lvbmFsIGRhIFVGTUcgcG9kZSwgc2VtIGFsdGVyYXIgbyBjb250Ze+/vWRvLCB0cmFuc3BvciBhIHN1YSBwdWJsaWNh77+977+9byBwYXJhIHF1YWxxdWVyIG1laW8gb3UgZm9ybWF0byBwYXJhIGZpbnMgZGUgcHJlc2VydmHvv73vv71vLgoKVm9j77+9IHRhbWLvv71tIGNvbmNvcmRhIHF1ZSBvIFJlcG9zaXTvv71yaW8gSW5zdGl0dWNpb25hbCBkYSBVRk1HIHBvZGUgbWFudGVyIG1haXMgZGUgdW1hIGPvv71waWEgZGUgc3VhIHB1YmxpY2Hvv73vv71vIHBhcmEgZmlucyBkZSBzZWd1cmFu77+9YSwgYmFjay11cCBlIHByZXNlcnZh77+977+9by4KClZvY++/vSBkZWNsYXJhIHF1ZSBhIHN1YSBwdWJsaWNh77+977+9byDvv70gb3JpZ2luYWwgZSBxdWUgdm9j77+9IHRlbSBvIHBvZGVyIGRlIGNvbmNlZGVyIG9zIGRpcmVpdG9zIGNvbnRpZG9zIG5lc3RhIGxpY2Vu77+9YS4gVm9j77+9IHRhbWLvv71tIGRlY2xhcmEgcXVlIG8gZGVw77+9c2l0byBkZSBzdWEgcHVibGljYe+/ve+/vW8gbu+/vW8sIHF1ZSBzZWphIGRlIHNldSBjb25oZWNpbWVudG8sIGluZnJpbmdlIGRpcmVpdG9zIGF1dG9yYWlzIGRlIG5pbmd177+9bS4KCkNhc28gYSBzdWEgcHVibGljYe+/ve+/vW8gY29udGVuaGEgbWF0ZXJpYWwgcXVlIHZvY++/vSBu77+9byBwb3NzdWkgYSB0aXR1bGFyaWRhZGUgZG9zIGRpcmVpdG9zIGF1dG9yYWlzLCB2b2Pvv70gZGVjbGFyYSBxdWUgb2J0ZXZlIGEgcGVybWlzc++/vW8gaXJyZXN0cml0YSBkbyBkZXRlbnRvciBkb3MgZGlyZWl0b3MgYXV0b3JhaXMgcGFyYSBjb25jZWRlciBhbyBSZXBvc2l077+9cmlvIEluc3RpdHVjaW9uYWwgZGEgVUZNRyBvcyBkaXJlaXRvcyBhcHJlc2VudGFkb3MgbmVzdGEgbGljZW7vv71hLCBlIHF1ZSBlc3NlIG1hdGVyaWFsIGRlIHByb3ByaWVkYWRlIGRlIHRlcmNlaXJvcyBlc3Tvv70gY2xhcmFtZW50ZSBpZGVudGlmaWNhZG8gZSByZWNvbmhlY2lkbyBubyB0ZXh0byBvdSBubyBjb250Ze+/vWRvIGRhIHB1YmxpY2Hvv73vv71vIG9yYSBkZXBvc2l0YWRhLgoKQ0FTTyBBIFBVQkxJQ0Hvv73vv71PIE9SQSBERVBPU0lUQURBIFRFTkhBIFNJRE8gUkVTVUxUQURPIERFIFVNIFBBVFJPQ++/vU5JTyBPVSBBUE9JTyBERSBVTUEgQUfvv71OQ0lBIERFIEZPTUVOVE8gT1UgT1VUUk8gT1JHQU5JU01PLCBWT0Pvv70gREVDTEFSQSBRVUUgUkVTUEVJVE9VIFRPRE9TIEUgUVVBSVNRVUVSIERJUkVJVE9TIERFIFJFVklT77+9TyBDT01PIFRBTULvv71NIEFTIERFTUFJUyBPQlJJR0Hvv73vv71FUyBFWElHSURBUyBQT1IgQ09OVFJBVE8gT1UgQUNPUkRPLgoKTyBSZXBvc2l077+9cmlvIEluc3RpdHVjaW9uYWwgZGEgVUZNRyBzZSBjb21wcm9tZXRlIGEgaWRlbnRpZmljYXIgY2xhcmFtZW50ZSBvIHNldSBub21lKHMpIG91IG8ocykgbm9tZXMocykgZG8ocykgZGV0ZW50b3IoZXMpIGRvcyBkaXJlaXRvcyBhdXRvcmFpcyBkYSBwdWJsaWNh77+977+9bywgZSBu77+9byBmYXLvv70gcXVhbHF1ZXIgYWx0ZXJh77+977+9bywgYWzvv71tIGRhcXVlbGFzIGNvbmNlZGlkYXMgcG9yIGVzdGEgbGljZW7vv71hLgo=Repositório de PublicaçõesPUBhttps://repositorio.ufmg.br/oaiopendoar:2022-02-18T21:47:14Repositório Institucional da UFMG - Universidade Federal de Minas Gerais (UFMG)false
dc.title.pt_BR.fl_str_mv PCR-RFLP screening of polymorphisms associated with benzimidazole resistance in Necator americanus and Ascaris lumbricoides from different geographical regions in Brazil
dc.title.alternative.pt_BR.fl_str_mv Triagem PCR-RFLP de polimorfismos associada à resistência ao benzimidazol em Necator americanus e Ascaris lumbricoides de diferentes regiões geográficas do Brasil
title PCR-RFLP screening of polymorphisms associated with benzimidazole resistance in Necator americanus and Ascaris lumbricoides from different geographical regions in Brazil
spellingShingle PCR-RFLP screening of polymorphisms associated with benzimidazole resistance in Necator americanus and Ascaris lumbricoides from different geographical regions in Brazil
Luciana Werneck Zuccherato
Ascaris lumbricoides
Necator americanus
title_short PCR-RFLP screening of polymorphisms associated with benzimidazole resistance in Necator americanus and Ascaris lumbricoides from different geographical regions in Brazil
title_full PCR-RFLP screening of polymorphisms associated with benzimidazole resistance in Necator americanus and Ascaris lumbricoides from different geographical regions in Brazil
title_fullStr PCR-RFLP screening of polymorphisms associated with benzimidazole resistance in Necator americanus and Ascaris lumbricoides from different geographical regions in Brazil
title_full_unstemmed PCR-RFLP screening of polymorphisms associated with benzimidazole resistance in Necator americanus and Ascaris lumbricoides from different geographical regions in Brazil
title_sort PCR-RFLP screening of polymorphisms associated with benzimidazole resistance in Necator americanus and Ascaris lumbricoides from different geographical regions in Brazil
author Luciana Werneck Zuccherato
author_facet Luciana Werneck Zuccherato
Luiz Fernando Vieira Furtado
Celi da Silva Medeiros
Carina da Silva Pinheiro
Elida Mara Leite Rabelo
author_role author
author2 Luiz Fernando Vieira Furtado
Celi da Silva Medeiros
Carina da Silva Pinheiro
Elida Mara Leite Rabelo
author2_role author
author
author
author
dc.contributor.author.fl_str_mv Luciana Werneck Zuccherato
Luiz Fernando Vieira Furtado
Celi da Silva Medeiros
Carina da Silva Pinheiro
Elida Mara Leite Rabelo
dc.subject.other.pt_BR.fl_str_mv Ascaris lumbricoides
Necator americanus
topic Ascaris lumbricoides
Necator americanus
description Ascaris lumbricoides and Necator americanus are soil-transmitted parasites with global geographic distribution, and they represent some of the most common and neglected infec tions in the world. Periodic treatment with mass drug administration (MDA) in endemic areas is the recommended action put forth by the World Health Organization. However, MDA can cause the selection of subpopulations that possess the genetic ability to overcome the mechanism of drug action. In fact, beta-tubulin gene mutations (codons 167, 198 and 200) are correlated with benzimidazole resistance in nematodes of veterinary importance. It is possible that these SNPs also have strong correlation with treatment resistance in the human geohelminths A. lumbricoides, Trichuris trichiura and hookworms. Here, we aimed to investigate the presence of some of these canonical molecular markers associated with par asite resistance to benzimidazole in N. americanus and A. lumbricoides collected from six Brazilian states. Nested-PCR and PCR-RFLP were used to detect mutations at codons 167 and 198 in 601 individual eggs of A. lumbricoides collected from 62 human stool samples; however, no mutations were found. Codons 198 and 200 were tested in 552 N. americanus eggs collected from 48 patients using the same methodology, which presented a relative fre quency of 1.4% and 1.1%, respectively. The presence of these SNPs in N. americanus eggs is an important finding, indicating that with high benzimidazole drug pressure there is potential for benzimidazole resistance to be selected in this hookworm. However, at these low frequencies it does not indicate that there is at present any benzimidazole resistance problem. This is the first systematic study performed in South America, and the study yielded a landscape of the genetic variants in the beta-tubulin gene and anthelmintic resis tance to soil-transmitted parasites detected by a simple, rapid and affordable genotyping assay of individual eggs
publishDate 2018
dc.date.issued.fl_str_mv 2018-09-17
dc.date.accessioned.fl_str_mv 2022-02-18T21:47:14Z
dc.date.available.fl_str_mv 2022-02-18T21:47:14Z
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.uri.fl_str_mv http://hdl.handle.net/1843/39486
dc.identifier.doi.pt_BR.fl_str_mv https://doi.org/10.1371/journal.pntd.0006766
dc.identifier.issn.pt_BR.fl_str_mv 03786978
url https://doi.org/10.1371/journal.pntd.0006766
http://hdl.handle.net/1843/39486
identifier_str_mv 03786978
dc.language.iso.fl_str_mv eng
language eng
dc.relation.ispartof.pt_BR.fl_str_mv Plos neglected tropical diseases
dc.rights.driver.fl_str_mv info:eu-repo/semantics/openAccess
eu_rights_str_mv openAccess
dc.format.none.fl_str_mv application/pdf
dc.publisher.none.fl_str_mv Universidade Federal de Minas Gerais
dc.publisher.initials.fl_str_mv UFMG
dc.publisher.country.fl_str_mv Brasil
dc.publisher.department.fl_str_mv ICB - INSTITUTO DE CIÊNCIAS BIOLOGICAS
publisher.none.fl_str_mv Universidade Federal de Minas Gerais
dc.source.none.fl_str_mv reponame:Repositório Institucional da UFMG
instname:Universidade Federal de Minas Gerais (UFMG)
instacron:UFMG
instname_str Universidade Federal de Minas Gerais (UFMG)
instacron_str UFMG
institution UFMG
reponame_str Repositório Institucional da UFMG
collection Repositório Institucional da UFMG
bitstream.url.fl_str_mv https://repositorio.ufmg.br/bitstream/1843/39486/1/License.txt
https://repositorio.ufmg.br/bitstream/1843/39486/2/PCR-RFLP_screening_of_polymorphisms_associated_wit.pdf
bitstream.checksum.fl_str_mv fa505098d172de0bc8864fc1287ffe22
734f26661c94c65e0760b0cc0f2bd6fa
bitstream.checksumAlgorithm.fl_str_mv MD5
MD5
repository.name.fl_str_mv Repositório Institucional da UFMG - Universidade Federal de Minas Gerais (UFMG)
repository.mail.fl_str_mv
_version_ 1797971107988373504