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

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. Su...

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Autores principales: Marcon, Helena Sanches, Coscrato, Virgínia Elias, Selivon, Denise, Perondini, André Luiz Paranhos, Marino, Celso Luis
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Sociedade Brasileira de Microbiologia 2011
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3769819/
https://www.ncbi.nlm.nih.gov/pubmed/24031693
http://dx.doi.org/10.1590/S1517-838220110002000046
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author Marcon, Helena Sanches
Coscrato, Virgínia Elias
Selivon, Denise
Perondini, André Luiz Paranhos
Marino, Celso Luis
author_facet Marcon, Helena Sanches
Coscrato, Virgínia Elias
Selivon, Denise
Perondini, André Luiz Paranhos
Marino, Celso Luis
author_sort Marcon, Helena Sanches
collection PubMed
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.
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spelling pubmed-37698192013-09-12 Variations in the sensitivity of different primers for detecting Wolbachia in Anastrepha (diptera: tephritidae) Marcon, Helena Sanches Coscrato, Virgínia Elias Selivon, Denise Perondini, André Luiz Paranhos Marino, Celso Luis Braz J Microbiol Genetics and Molecular Microbiology 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. Sociedade Brasileira de Microbiologia 2011 2011-06-01 /pmc/articles/PMC3769819/ /pubmed/24031693 http://dx.doi.org/10.1590/S1517-838220110002000046 Text en © Sociedade Brasileira de Microbiologia http://creativecommons.org/licenses/by-nc/3.0/ All the content of the journal, except where otherwise noted, is licensed under a Creative Commons License
spellingShingle Genetics and Molecular Microbiology
Marcon, Helena Sanches
Coscrato, Virgínia Elias
Selivon, Denise
Perondini, André Luiz Paranhos
Marino, Celso Luis
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)
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_short 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)
topic Genetics and Molecular Microbiology
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3769819/
https://www.ncbi.nlm.nih.gov/pubmed/24031693
http://dx.doi.org/10.1590/S1517-838220110002000046
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