In-Field Capable Loop-Mediated Isothermal Amplification Detection of Spodoptera frugiperda (Lepidoptera: Noctuidae) Larvae Using a Rapid and Simple Crude Extraction Technique

Fall armyworm, Spodoptera frugiperda J.E. Smith (Lepidoptera: Noctuidae), is an economically important pest worldwide and has recently been identified in Australia. Morphological identification of S. frugiperda at early larval stages can be difficult often requiring expert microscopy analysis. Rapid...

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Bibliographic Details
Published in:Journal of economic entomology 2021-12, Vol.114 (6), p.2610-2614
Main Authors: Congdon, B. S, Webster, C. G, Severtson, D, Spafford, H
Format: Article
Language:English
Subjects:
Analysis
Animals
Deoxyribonucleic acid
DNA
Economic importance
fall armyworm
Gene amplification
Larva - genetics
Larvae
Lepidoptera
Methods
molecular diagnostic
Molecular Diagnostic Techniques
Moths
Noctuidae
Nucleic Acid Amplification Techniques
Oxidases
pest management
SHORT COMMUNICATION
Spodoptera - genetics
Spodoptera frugiperda
Zea mays
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Summary:Fall armyworm, Spodoptera frugiperda J.E. Smith (Lepidoptera: Noctuidae), is an economically important pest worldwide and has recently been identified in Australia. Morphological identification of S. frugiperda at early larval stages can be difficult often requiring expert microscopy analysis. Rapid and accurate in-field diagnosis is vital for management decision support and there are no tools currently available for this purpose. In this study, a sensitive, specific, and in-field capable loop-mediated isothermal amplification (LAMP) assay was developed to detect S. frugiperda larvae. A primer set based on a highly conserved region of the S. frugiperda cytochrome oxidase subunit 1 (COX1) gene provided detection within 30 min from both total DNA and crude extractions. The crude extraction technique of crushing 10 mg of S. frugiperda material in 50 µl ddH2O and further diluting the homogenate in ddH2O is rapid, simple, and does not require heat blocks, centrifuges, or special buffers increasing its utility as a field-based technique. The primer set detected as little as 24 pg of S. frugiperda DNA and did not cross-react with any other of the lepidopteran species tested that are easily confused with S. frugiperda in Australia. Therefore, this assay could be used in-field to correctly identify the presence of S. frugiperda and thereby greatly assist with timely management decisions.
ISSN:0022-0493
1938-291X
DOI:10.1093/jee/toab168