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Development of a simple and accurate molecular tool for Spodoptera frugiperda species identification using LAMP
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  • Juil KIM,
  • Hwa Yeun Nam,
  • Min Kwon,
  • Hyun Ju Kim,
  • Hwi-Jong Yi,
  • Sabine Hänniger,
  • Melanie Unbehend,
  • David Heckel
Juil KIM
National Institute of Crop Science
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Hwa Yeun Nam
National Institute of Crop Science
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Min Kwon
National Institute of Crop Science
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Hyun Ju Kim
National Institute of Crop Science
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Hwi-Jong Yi
National Institute of Crop Science Department of Functional Crop
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Sabine Hänniger
Max Planck Institute for Chemical Ecology
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Melanie Unbehend
Max-Planck-Institute for Chemical Ecology
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David Heckel
Max Planck Institute for Chemical Ecology
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Abstract

The fall armyworm, Spodoptera frugiperda is a native species in the Americas. However, nowadays it is one of the most serious invasive lepidopteran pests in African and Asian countries. S. frugiperda has been spread very quickly after the first outbreak was reported in many countries. Based on mt genome sequence alignment, S. frugiperda specific sequence region was identified in tRNAs coding region between NADH dehydrogenase, ND3 and ND5. By using this unique region, species diagnostic primers were designed and applied in LAMP (lamp loop mediated isothermal amplification) assay as well as conventional PCR to identify the field-collected samples of S. frugiperda. Optimal incubation condition of LAMP assay was 61℃ for 90 minutes with 4 LAMP primers, and additional loop primer increased the amplification efficiency. Also, wide range of DNA concentration responded in LAMP assay and minimum detectable DNA concentration was 10 pg. This LAMP assay was also applied in DNA releasing technique from larval and adult sample, without DNA extraction, 95℃ incubation for five minutes of the tissue sample. This new molecular diagnostic method is easy to use and accurate. It possibly applied in intensive field monitoring of S. frugiperda and its ecological studies.