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Quantification of the Pirimicarb Resistance Allele Frequency in Pooled Cotton Aphid (Aphis gossypii Glover) Samples by TaqMan SNP Genotyping Assay
BACKGROUND: Pesticide resistance monitoring is a crucial part to achieving sustainable integrated pest management (IPM) in agricultural production systems. Monitoring of resistance in arthropod populations is initially performed by bioassay, a method that detects a phenotypic response to pesticides....
Autores principales: | , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Public Library of Science
2014
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3948748/ https://www.ncbi.nlm.nih.gov/pubmed/24614533 http://dx.doi.org/10.1371/journal.pone.0091104 |
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author | Chen, Yizhou Bogema, Daniel R. Barchia, Idris M. Herron, Grant A. |
author_facet | Chen, Yizhou Bogema, Daniel R. Barchia, Idris M. Herron, Grant A. |
author_sort | Chen, Yizhou |
collection | PubMed |
description | BACKGROUND: Pesticide resistance monitoring is a crucial part to achieving sustainable integrated pest management (IPM) in agricultural production systems. Monitoring of resistance in arthropod populations is initially performed by bioassay, a method that detects a phenotypic response to pesticides. Molecular diagnostic assays, offering speed and cost improvements, can be developed when the causative mutation for resistance has been identified. However, improvements to throughput are limited as genotyping methods cannot be accurately applied to pooled DNA. Quantifying an allele frequency from pooled DNA would allow faster and cheaper monitoring of pesticide resistance. METHODOLOGY/PRINCIPAL FINDINGS: We demonstrate a new method to quantify a resistance allele frequency (RAF) from pooled insects via TaqMan assay by using raw fluorescence data to calculate the transformed fluorescence ratio k’ at the inflexion point based on a four parameter sigmoid curve. Our results show that k’ is reproducible and highly correlated with RAF (r >0.99). We also demonstrate that k’ has a non-linear relationship with RAF and that five standard points are sufficient to build a prediction model. Additionally, we identified a non-linear relationship between runs for k’, allowing the combination of samples across multiple runs in a single analysis. CONCLUSIONS/SIGNIFICANCE: The transformed fluorescence ratio (k') method can be used to monitor pesticide resistance in IPM and to accurately quantify allele frequency from pooled samples. We have determined that five standards (0.0, 0.2, 0.5, 0.8, and 1.0) are sufficient for accurate prediction and are statistically-equivalent to the 13 standard points used experimentally |
format | Online Article Text |
id | pubmed-3948748 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2014 |
publisher | Public Library of Science |
record_format | MEDLINE/PubMed |
spelling | pubmed-39487482014-03-13 Quantification of the Pirimicarb Resistance Allele Frequency in Pooled Cotton Aphid (Aphis gossypii Glover) Samples by TaqMan SNP Genotyping Assay Chen, Yizhou Bogema, Daniel R. Barchia, Idris M. Herron, Grant A. PLoS One Research Article BACKGROUND: Pesticide resistance monitoring is a crucial part to achieving sustainable integrated pest management (IPM) in agricultural production systems. Monitoring of resistance in arthropod populations is initially performed by bioassay, a method that detects a phenotypic response to pesticides. Molecular diagnostic assays, offering speed and cost improvements, can be developed when the causative mutation for resistance has been identified. However, improvements to throughput are limited as genotyping methods cannot be accurately applied to pooled DNA. Quantifying an allele frequency from pooled DNA would allow faster and cheaper monitoring of pesticide resistance. METHODOLOGY/PRINCIPAL FINDINGS: We demonstrate a new method to quantify a resistance allele frequency (RAF) from pooled insects via TaqMan assay by using raw fluorescence data to calculate the transformed fluorescence ratio k’ at the inflexion point based on a four parameter sigmoid curve. Our results show that k’ is reproducible and highly correlated with RAF (r >0.99). We also demonstrate that k’ has a non-linear relationship with RAF and that five standard points are sufficient to build a prediction model. Additionally, we identified a non-linear relationship between runs for k’, allowing the combination of samples across multiple runs in a single analysis. CONCLUSIONS/SIGNIFICANCE: The transformed fluorescence ratio (k') method can be used to monitor pesticide resistance in IPM and to accurately quantify allele frequency from pooled samples. We have determined that five standards (0.0, 0.2, 0.5, 0.8, and 1.0) are sufficient for accurate prediction and are statistically-equivalent to the 13 standard points used experimentally Public Library of Science 2014-03-10 /pmc/articles/PMC3948748/ /pubmed/24614533 http://dx.doi.org/10.1371/journal.pone.0091104 Text en © 2014 Chen et al http://creativecommons.org/licenses/by/4.0/ This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are properly credited. |
spellingShingle | Research Article Chen, Yizhou Bogema, Daniel R. Barchia, Idris M. Herron, Grant A. Quantification of the Pirimicarb Resistance Allele Frequency in Pooled Cotton Aphid (Aphis gossypii Glover) Samples by TaqMan SNP Genotyping Assay |
title | Quantification of the Pirimicarb Resistance Allele Frequency in Pooled Cotton Aphid (Aphis gossypii Glover) Samples by TaqMan SNP Genotyping Assay |
title_full | Quantification of the Pirimicarb Resistance Allele Frequency in Pooled Cotton Aphid (Aphis gossypii Glover) Samples by TaqMan SNP Genotyping Assay |
title_fullStr | Quantification of the Pirimicarb Resistance Allele Frequency in Pooled Cotton Aphid (Aphis gossypii Glover) Samples by TaqMan SNP Genotyping Assay |
title_full_unstemmed | Quantification of the Pirimicarb Resistance Allele Frequency in Pooled Cotton Aphid (Aphis gossypii Glover) Samples by TaqMan SNP Genotyping Assay |
title_short | Quantification of the Pirimicarb Resistance Allele Frequency in Pooled Cotton Aphid (Aphis gossypii Glover) Samples by TaqMan SNP Genotyping Assay |
title_sort | quantification of the pirimicarb resistance allele frequency in pooled cotton aphid (aphis gossypii glover) samples by taqman snp genotyping assay |
topic | Research Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3948748/ https://www.ncbi.nlm.nih.gov/pubmed/24614533 http://dx.doi.org/10.1371/journal.pone.0091104 |
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