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Multiple TaqMan qPCR and droplet digital PCR (ddPCR) diagnostics for pesticide resistance monitoring and management, in the major agricultural pest Tetranychus urticae

(2022) PEST MANAGEMENT SCIENCE. 78(1). p.263-273
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  • SuperPests (Innovative tools for rational control of the most difficult-to-manage pests (super pests) and the diseases they transmit)
Abstract
BACKGROUND Decisions on which pesticide to use in agriculture are expected to become more difficult, as the number of available chemicals is decreasing. For Tetranychus urticae (T. urticae), a major pest for which a number of candidate markers for pesticide resistance are in place, molecular diagnostics could support decision-making for the rational use of acaricides. RESULTS A suite of 12 TaqMan qPCR assays [G314D (GluCl1), G326E, I321T (GluCl3), G119S, F331W (Ace-1), H92R (PSST), L1024V, F1538I (VGSC), I1017F (CHS1), G126S, S141F, P262T (cytb)], were validated against Sanger-sequencing, and subsequently adapted for use with the ddPCR technology. The concordance correlation coefficient between the actual and ddPCR measured mutant allelic frequencies was 0.995 (95% CI = 0.991-0.998), and no systematic, proportional, or random differences were detected. The achieved Limit of Detection (LoD) was 0.1% (detection of one mutant in a background of 999 wild type mites). The ddPCR assay panel was then assessed in terms of agreement with phenotypic resistance, through a pilot application in field populations from Crete, with strong correlation and thus predictive and diagnostic value of the molecular assays in some cases (e.g., etoxazole and abamectin resistance). Molecular diagnostics were able to capture incipient resistance that was otherwise missed by phenotypic bioassays. The molecular and phenotypic resistance screening of T. urticae field populations from Crete, revealed both multi-resistant and susceptible populations. CONCLUSION The highly sensitive T. urticae molecular diagnostic platforms developed in this study could prove a valuable tool for pesticide resistance management.
Keywords
2-SPOTTED SPIDER-MITE, TARGET-SITE, INSECTICIDE RESISTANCE, ABAMECTIN, RESISTANCE, CROSS-RESISTANCE, KOCH ACARI, MOLECULAR DIAGNOSTICS, MUTATIONS, EVOLUTION, CHANNEL, ddPCR, TaqMan diagnostics, agricultural pests, insecticide resistance, management, target-site resistance

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MLA
Mavridis, Konstantinos, et al. “Multiple TaqMan QPCR and Droplet Digital PCR (DdPCR) Diagnostics for Pesticide Resistance Monitoring and Management, in the Major Agricultural Pest Tetranychus Urticae.” PEST MANAGEMENT SCIENCE, vol. 78, no. 1, 2022, pp. 263–73, doi:10.1002/ps.6632.
APA
Mavridis, K., Papapostolou, K. M., Riga, M., Ilias, A., Michaelidou, K., Bass, C., … Vontas, J. (2022). Multiple TaqMan qPCR and droplet digital PCR (ddPCR) diagnostics for pesticide resistance monitoring and management, in the major agricultural pest Tetranychus urticae. PEST MANAGEMENT SCIENCE, 78(1), 263–273. https://doi.org/10.1002/ps.6632
Chicago author-date
Mavridis, Konstantinos, Kyriaki Maria Papapostolou, Maria Riga, Aris Ilias, Kleita Michaelidou, Chris Bass, Thomas Van Leeuwen, Anastasia Tsagkarakou, and John Vontas. 2022. “Multiple TaqMan QPCR and Droplet Digital PCR (DdPCR) Diagnostics for Pesticide Resistance Monitoring and Management, in the Major Agricultural Pest Tetranychus Urticae.” PEST MANAGEMENT SCIENCE 78 (1): 263–73. https://doi.org/10.1002/ps.6632.
Chicago author-date (all authors)
Mavridis, Konstantinos, Kyriaki Maria Papapostolou, Maria Riga, Aris Ilias, Kleita Michaelidou, Chris Bass, Thomas Van Leeuwen, Anastasia Tsagkarakou, and John Vontas. 2022. “Multiple TaqMan QPCR and Droplet Digital PCR (DdPCR) Diagnostics for Pesticide Resistance Monitoring and Management, in the Major Agricultural Pest Tetranychus Urticae.” PEST MANAGEMENT SCIENCE 78 (1): 263–273. doi:10.1002/ps.6632.
Vancouver
1.
Mavridis K, Papapostolou KM, Riga M, Ilias A, Michaelidou K, Bass C, et al. Multiple TaqMan qPCR and droplet digital PCR (ddPCR) diagnostics for pesticide resistance monitoring and management, in the major agricultural pest Tetranychus urticae. PEST MANAGEMENT SCIENCE. 2022;78(1):263–73.
IEEE
[1]
K. Mavridis et al., “Multiple TaqMan qPCR and droplet digital PCR (ddPCR) diagnostics for pesticide resistance monitoring and management, in the major agricultural pest Tetranychus urticae,” PEST MANAGEMENT SCIENCE, vol. 78, no. 1, pp. 263–273, 2022.
@article{8746272,
  abstract     = {{BACKGROUND Decisions on which pesticide to use in agriculture are expected to become more difficult, as the number of available chemicals is decreasing. For Tetranychus urticae (T. urticae), a major pest for which a number of candidate markers for pesticide resistance are in place, molecular diagnostics could support decision-making for the rational use of acaricides. RESULTS A suite of 12 TaqMan qPCR assays [G314D (GluCl1), G326E, I321T (GluCl3), G119S, F331W (Ace-1), H92R (PSST), L1024V, F1538I (VGSC), I1017F (CHS1), G126S, S141F, P262T (cytb)], were validated against Sanger-sequencing, and subsequently adapted for use with the ddPCR technology. The concordance correlation coefficient between the actual and ddPCR measured mutant allelic frequencies was 0.995 (95% CI = 0.991-0.998), and no systematic, proportional, or random differences were detected. The achieved Limit of Detection (LoD) was 0.1% (detection of one mutant in a background of 999 wild type mites). The ddPCR assay panel was then assessed in terms of agreement with phenotypic resistance, through a pilot application in field populations from Crete, with strong correlation and thus predictive and diagnostic value of the molecular assays in some cases (e.g., etoxazole and abamectin resistance). Molecular diagnostics were able to capture incipient resistance that was otherwise missed by phenotypic bioassays. The molecular and phenotypic resistance screening of T. urticae field populations from Crete, revealed both multi-resistant and susceptible populations. CONCLUSION The highly sensitive T. urticae molecular diagnostic platforms developed in this study could prove a valuable tool for pesticide resistance management.}},
  author       = {{Mavridis, Konstantinos and Papapostolou, Kyriaki Maria and Riga, Maria and Ilias, Aris and Michaelidou, Kleita and Bass, Chris and Van Leeuwen, Thomas and Tsagkarakou, Anastasia and Vontas, John}},
  issn         = {{1526-498X}},
  journal      = {{PEST MANAGEMENT SCIENCE}},
  keywords     = {{2-SPOTTED SPIDER-MITE,TARGET-SITE,INSECTICIDE RESISTANCE,ABAMECTIN,RESISTANCE,CROSS-RESISTANCE,KOCH ACARI,MOLECULAR DIAGNOSTICS,MUTATIONS,EVOLUTION,CHANNEL,ddPCR,TaqMan diagnostics,agricultural pests,insecticide resistance,management,target-site resistance}},
  language     = {{eng}},
  number       = {{1}},
  pages        = {{263--273}},
  title        = {{Multiple TaqMan qPCR and droplet digital PCR (ddPCR) diagnostics for pesticide resistance monitoring and management, in the major agricultural pest Tetranychus urticae}},
  url          = {{http://doi.org/10.1002/ps.6632}},
  volume       = {{78}},
  year         = {{2022}},
}

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