Variations in the sensitivity of different primers for detecting Wolbachia in Anastrepha (diptera: tephritidae)

Detalhes bibliográficos
Autor(a) principal: Marcon, Helena Sanches [UNESP]
Data de Publicação: 2011
Outros Autores: Coscrato, Virgínia Elias [UNESP], Selivon, Denise, Perondini, André Luiz Paranhos, Marino, Celso Luis [UNESP]
Tipo de documento: Artigo
Idioma: eng
Título da fonte: Repositório Institucional da UNESP
Texto Completo: http://dx.doi.org/10.1590/S1517-83822011000200046
http://hdl.handle.net/11449/26644
Resumo: Wolbachia are endosymbiont bacteria of the family Rickettsiacea that are widespread in invertebrates and occur between 20% and 60% of Neotropical insects. These bacteria are responsible for reproductive phenomena such as cytoplasmic incompatibility, male killing, feminization and parthenogenesis. Supergroups A and B of Wolbachia are common in insects and can be identified using primers for 16S rDNA, ftsZ and wsp; these primers vary in their ability to detect Wolbachia. The ftsZ primer was the first primer used to detect Wolbachia in Anastrepha fruit flies. The primers for 16S rDNA, ftsZ and wsp and the corresponding PCR conditions have been optimized to study the distribution of Wolbachia and their effect on the biology of Anastrepha in Brazil. In this work, we examined the ability of these primers to detect Wolbachia in Anastrepha populations from three regions in the State of São Paulo, southeastern Brazil. All of the samples were positive for Wolbachia supergroup A when screened with primers for 16S A rDNA and wsp A; the wsp B primer also gave a positive result, indicating cross-reactivity. The ftsZ primer showed a poor ability to detect Wolbachia in Anastrepha and generated false negatives in 44.9% of the samples. These findings indicate that reliable PCR detection of Wolbachia requires the use of primers for 16S rDNA and wsp to avoid cross-reactions and false negatives, and that the ftsZ primer needs to be redesigned to improve its selectivity.
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spelling Variations in the sensitivity of different primers for detecting Wolbachia in Anastrepha (diptera: tephritidae)Anastrephacross reactionftsZwspWolbachia are endosymbiont bacteria of the family Rickettsiacea that are widespread in invertebrates and occur between 20% and 60% of Neotropical insects. These bacteria are responsible for reproductive phenomena such as cytoplasmic incompatibility, male killing, feminization and parthenogenesis. Supergroups A and B of Wolbachia are common in insects and can be identified using primers for 16S rDNA, ftsZ and wsp; these primers vary in their ability to detect Wolbachia. The ftsZ primer was the first primer used to detect Wolbachia in Anastrepha fruit flies. The primers for 16S rDNA, ftsZ and wsp and the corresponding PCR conditions have been optimized to study the distribution of Wolbachia and their effect on the biology of Anastrepha in Brazil. In this work, we examined the ability of these primers to detect Wolbachia in Anastrepha populations from three regions in the State of São Paulo, southeastern Brazil. All of the samples were positive for Wolbachia supergroup A when screened with primers for 16S A rDNA and wsp A; the wsp B primer also gave a positive result, indicating cross-reactivity. The ftsZ primer showed a poor ability to detect Wolbachia in Anastrepha and generated false negatives in 44.9% of the samples. These findings indicate that reliable PCR detection of Wolbachia requires the use of primers for 16S rDNA and wsp to avoid cross-reactions and false negatives, and that the ftsZ primer needs to be redesigned to improve its selectivity.Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)Universidade Estadual Paulista Júlio de Mesquita Filho Instituto de Biociências Departamento de GenéticaUniversidade de São Paulo Instituto de Biociências Departamento de Genética e Biologia EvolutivaUniversidade Estadual Paulista Júlio de Mesquita Filho Instituto de Biociências Departamento de GenéticaSociedade Brasileira de MicrobiologiaUniversidade Estadual Paulista (Unesp)Universidade de São Paulo (USP)Marcon, Helena Sanches [UNESP]Coscrato, Virgínia Elias [UNESP]Selivon, DenisePerondini, André Luiz ParanhosMarino, Celso Luis [UNESP]2014-05-20T15:07:44Z2014-05-20T15:07:44Z2011-06-01info:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/article778-785application/pdfhttp://dx.doi.org/10.1590/S1517-83822011000200046Brazilian Journal of Microbiology. Sociedade Brasileira de Microbiologia, v. 42, n. 2, p. 778-785, 2011.1517-8382http://hdl.handle.net/11449/2664410.1590/S1517-83822011000200046S1517-83822011000200046WOS:000291614000046S1517-83822011000200046.pdf01653487382083190000-0003-4524-954XSciELOreponame:Repositório Institucional da UNESPinstname:Universidade Estadual Paulista (UNESP)instacron:UNESPengBrazilian Journal of Microbiology1.8100,630info:eu-repo/semantics/openAccess2023-11-07T06:11:35Zoai:repositorio.unesp.br:11449/26644Repositório InstitucionalPUBhttp://repositorio.unesp.br/oai/requestopendoar:29462024-08-05T17:05:05.847221Repositório Institucional da UNESP - Universidade Estadual Paulista (UNESP)false
dc.title.none.fl_str_mv Variations in the sensitivity of different primers for detecting Wolbachia in Anastrepha (diptera: tephritidae)
title Variations in the sensitivity of different primers for detecting Wolbachia in Anastrepha (diptera: tephritidae)
spellingShingle Variations in the sensitivity of different primers for detecting Wolbachia in Anastrepha (diptera: tephritidae)
Marcon, Helena Sanches [UNESP]
Anastrepha
cross reaction
ftsZ
wsp
title_short Variations in the sensitivity of different primers for detecting Wolbachia in Anastrepha (diptera: tephritidae)
title_full Variations in the sensitivity of different primers for detecting Wolbachia in Anastrepha (diptera: tephritidae)
title_fullStr Variations in the sensitivity of different primers for detecting Wolbachia in Anastrepha (diptera: tephritidae)
title_full_unstemmed Variations in the sensitivity of different primers for detecting Wolbachia in Anastrepha (diptera: tephritidae)
title_sort Variations in the sensitivity of different primers for detecting Wolbachia in Anastrepha (diptera: tephritidae)
author Marcon, Helena Sanches [UNESP]
author_facet Marcon, Helena Sanches [UNESP]
Coscrato, Virgínia Elias [UNESP]
Selivon, Denise
Perondini, André Luiz Paranhos
Marino, Celso Luis [UNESP]
author_role author
author2 Coscrato, Virgínia Elias [UNESP]
Selivon, Denise
Perondini, André Luiz Paranhos
Marino, Celso Luis [UNESP]
author2_role author
author
author
author
dc.contributor.none.fl_str_mv Universidade Estadual Paulista (Unesp)
Universidade de São Paulo (USP)
dc.contributor.author.fl_str_mv Marcon, Helena Sanches [UNESP]
Coscrato, Virgínia Elias [UNESP]
Selivon, Denise
Perondini, André Luiz Paranhos
Marino, Celso Luis [UNESP]
dc.subject.por.fl_str_mv Anastrepha
cross reaction
ftsZ
wsp
topic Anastrepha
cross reaction
ftsZ
wsp
description Wolbachia are endosymbiont bacteria of the family Rickettsiacea that are widespread in invertebrates and occur between 20% and 60% of Neotropical insects. These bacteria are responsible for reproductive phenomena such as cytoplasmic incompatibility, male killing, feminization and parthenogenesis. Supergroups A and B of Wolbachia are common in insects and can be identified using primers for 16S rDNA, ftsZ and wsp; these primers vary in their ability to detect Wolbachia. The ftsZ primer was the first primer used to detect Wolbachia in Anastrepha fruit flies. The primers for 16S rDNA, ftsZ and wsp and the corresponding PCR conditions have been optimized to study the distribution of Wolbachia and their effect on the biology of Anastrepha in Brazil. In this work, we examined the ability of these primers to detect Wolbachia in Anastrepha populations from three regions in the State of São Paulo, southeastern Brazil. All of the samples were positive for Wolbachia supergroup A when screened with primers for 16S A rDNA and wsp A; the wsp B primer also gave a positive result, indicating cross-reactivity. The ftsZ primer showed a poor ability to detect Wolbachia in Anastrepha and generated false negatives in 44.9% of the samples. These findings indicate that reliable PCR detection of Wolbachia requires the use of primers for 16S rDNA and wsp to avoid cross-reactions and false negatives, and that the ftsZ primer needs to be redesigned to improve its selectivity.
publishDate 2011
dc.date.none.fl_str_mv 2011-06-01
2014-05-20T15:07:44Z
2014-05-20T15:07:44Z
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://dx.doi.org/10.1590/S1517-83822011000200046
Brazilian Journal of Microbiology. Sociedade Brasileira de Microbiologia, v. 42, n. 2, p. 778-785, 2011.
1517-8382
http://hdl.handle.net/11449/26644
10.1590/S1517-83822011000200046
S1517-83822011000200046
WOS:000291614000046
S1517-83822011000200046.pdf
0165348738208319
0000-0003-4524-954X
url http://dx.doi.org/10.1590/S1517-83822011000200046
http://hdl.handle.net/11449/26644
identifier_str_mv Brazilian Journal of Microbiology. Sociedade Brasileira de Microbiologia, v. 42, n. 2, p. 778-785, 2011.
1517-8382
10.1590/S1517-83822011000200046
S1517-83822011000200046
WOS:000291614000046
S1517-83822011000200046.pdf
0165348738208319
0000-0003-4524-954X
dc.language.iso.fl_str_mv eng
language eng
dc.relation.none.fl_str_mv Brazilian Journal of Microbiology
1.810
0,630
dc.rights.driver.fl_str_mv info:eu-repo/semantics/openAccess
eu_rights_str_mv openAccess
dc.format.none.fl_str_mv 778-785
application/pdf
dc.publisher.none.fl_str_mv Sociedade Brasileira de Microbiologia
publisher.none.fl_str_mv Sociedade Brasileira de Microbiologia
dc.source.none.fl_str_mv SciELO
reponame:Repositório Institucional da UNESP
instname:Universidade Estadual Paulista (UNESP)
instacron:UNESP
instname_str Universidade Estadual Paulista (UNESP)
instacron_str UNESP
institution UNESP
reponame_str Repositório Institucional da UNESP
collection Repositório Institucional da UNESP
repository.name.fl_str_mv Repositório Institucional da UNESP - Universidade Estadual Paulista (UNESP)
repository.mail.fl_str_mv
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