Development of a Rapid Isothermal Amplification Assay for the Fall Armyworm, <i>Spodoptera frugiperda</i> (Lepidoptera: Noctuidae), Using Species-Specific Genomic Sequences
Jeong Sun Park,
Keon Hee Lee,
Min Jee Kim,
Deuk-Soo Choi,
Kyeong-Yeoll Lee,
Tariku Tesfaye Edosa,
Teshale Daba Dinka,
Woori Kwak,
Iksoo Kim
Affiliations
Jeong Sun Park
Department of Applied Biology, College of Agriculture and Life Sciences, Chonnam National University, Gwangju 61186, Republic of Korea
Keon Hee Lee
Department of Applied Biology, College of Agriculture and Life Sciences, Chonnam National University, Gwangju 61186, Republic of Korea
Min Jee Kim
Experiment and Analysis Division, Honam Regional Office, Animal and Plant Quarantine Agency, Gunsan 54096, Republic of Korea
Deuk-Soo Choi
Quarantine Technology Institute Ins., Gimcheon 39660, Republic of Korea
Kyeong-Yeoll Lee
Department of Applied Biosciences, College of Agriculture and Life Sciences, Kyungpook National University, Daegu 41566, Republic of Korea
Tariku Tesfaye Edosa
Ethiopian Institute of Agricultural Research, Ambo Agricultural Research Center, Ambo P.O. Box 37, Ethiopia
Teshale Daba Dinka
Ethiopian Institute of Agricultural Research, Ambo Agricultural Research Center, Ambo P.O. Box 37, Ethiopia
Woori Kwak
Department of Medical and Biological Sciences, The Catholic University of Korea, Bucheon 14662, Republic of Korea
Iksoo Kim
Department of Applied Biology, College of Agriculture and Life Sciences, Chonnam National University, Gwangju 61186, Republic of Korea
The fall armyworm (FAW), Spodoptera frugiperda (Lepidoptera: Noctuidae), is native to tropical and subtropical regions of the Western Hemisphere, but is now regularly appearing in crop fields across South Korea, particularly in corn fields. Therefore, it is crucial to promptly and accurately identify the presence of FAW in crop fields to effectively eradicate it as a regulated quarantine species. We developed a loop-mediated isothermal amplification (LAMP) assay, which allows for rapid in-filed identification. To develop the LAMP assay, we selected FAW-specific genomic regions from the whole-genome sequences of one FAW and 13 other lepidopteran species and validated five primer sets that consistently produced positive reactions in ten FAW samples collected from eight different locations in four countries. The assay successfully identified FAW in a maximum of 45 min, starting from crude DNA extraction (~15 min) to diagnosis (30 min) from the following samples, which were deposited outdoors for 30 days: a 1st-instar larva, an adult leg, an adult antenna, and 1/16 and 1/8 of an adult thorax. The five assays can be used selectively or in combination to cross-check and provide further confidence in the in-field diagnosis of FAW.
