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A diagnostic LAMP assay for rapid identification of an invasive plant pest, fall armyworm Spodoptera frugiperda (Lepidoptera: Noctuidae)

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Agarwal, A., Rako, L., Schutze, M. K., Starkie, M. L., Tay, W. T., Rodoni, B. C. and Blacket, M. J. (2022) A diagnostic LAMP assay for rapid identification of an invasive plant pest, fall armyworm Spodoptera frugiperda (Lepidoptera: Noctuidae). Scientific Reports, 12 (1). p. 1116. ISSN 2045-2322

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Article Link: https://doi.org/10.1038/s41598-021-04496-x

Abstract

Fall armyworm (FAW), Spodoptera frugiperda (Lepidoptera: Noctuidae), is a highly polyphagous invasive plant pest that has expanded its global geographic distribution, including recently into much of Australia. Rapid diagnostic tests are required for identification of FAW to assist subsequent management and control. We developed a new loop-mediated isothermal amplification (LAMP) assay based on the mitochondrial cytochrome c oxidase subunit I (COI) gene for accurate and timely diagnosis of FAW in the field. The specificity of the new assay was tested against a broad panel of twenty non-target noctuids, including eight other Spodoptera species. Only S. frugiperda samples produced amplification within 20 min, with an anneal derivative temperature of 78.3 ± 0.3 °C. A gBlock dsDNA fragment was developed and trialled as a synthetic positive control, with a different anneal derivative of 81 °C. The new FAW LAMP assay was able to detect FAW DNA down to 2.4 pg, similar to an existing laboratory-based real-time PCR assay. We also trialled the new FAW assay with a colorimetric master mix and found it could successfully amplify positive FAW samples in half the time compared to an existing FAW colorimetric LAMP assay. Given the high sensitivity and rapid amplification time, we recommend the use of this newly developed FAW LAMP assay in a portable real-time fluorometer for in-field diagnosis of FAW.

Item Type:Article
Business groups:Biosecurity Queensland
Additional Information:Open access
Subjects:Science > Entomology
Science > Invasive Species > Animals
Plant culture > Field crops
Plant pests and diseases > Plant pathology
Plant pests and diseases > Pest control and treatment of diseases. Plant protection
Live Archive:27 Jan 2022 00:38
Last Modified:27 Jan 2022 00:38

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