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6 April 2023 Field-based recombinase polymerase amplification and lab-based qPCR assays for detection of Helicoverpa armigera
Mitchell Rich, Enoch Noh, Hehe Wang, Jeremy Greene, Todd Gilligan, Francis P.F. Reay-Jones, Matt Turnbull, Frida Zink
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Abstract

Helicoverpa armigera (Hübner) is a major crop pest native to Europe, Asia, Australia, and Africa which has recently invaded South America and has caused billions of dollars in agricultural losses. Because of challenges in differentiating between H. armigera and Helicoverpa zea (Boddie), a closely related species native to North and South America, genetic tests have previously been developed to detect H. armigera DNA in pooled samples of moth legs. In this study, a field-based recombinase polymerase amplification (RPA) assay using a lateral flow strip and a qPCR melt curve assay were developed for specific detection of H. armigera DNA in pooled moth samples. In addition, a crude DNA extraction protocol for whole moths was developed to allow rapid preparation of DNA samples.The RPA field test was able to detect ≥ 10 pg of purified H. armigera DNA and the crude DNA of one H. armigera sample in a background of 999 H. zea equivalents. The qPCR assay was able to detect ≥ 100 fg of purified H. armigera DNA and the crude DNA of one H. armigera sample in a background of up to 99,999 H. zea equivalents. Both RPA and qPCR assays detected H. armigera in the crude DNA extracted in the field from a pool of one H. armigera moth and 999 H. zea moths. These newly developed molecular assays to detect H. armigera will contribute to large-scale surveillance programs of H. armigera.

Mitchell Rich, Enoch Noh, Hehe Wang, Jeremy Greene, Todd Gilligan, Francis P.F. Reay-Jones, Matt Turnbull, and Frida Zink "Field-based recombinase polymerase amplification and lab-based qPCR assays for detection of Helicoverpa armigera," Journal of Economic Entomology 116(3), 973-982, (6 April 2023). https://doi.org/10.1093/jee/toad048
Received: 7 November 2022; Accepted: 6 March 2023; Published: 6 April 2023
JOURNAL ARTICLE
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KEYWORDS
Helicoverpa zea
old world bollworm
qPCR
RPA
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