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A Rapid and Cost-Effective Identification of Invertebrate Pests at the Borders Using MinION Sequencing of DNA Barcodes

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Abeynayake, S. W., Fiorito, S., Dinsdale, A., Whattam, M., Crowe, B., Sparks, K., Campbell, P. R. and Gambley, C. (2021) A Rapid and Cost-Effective Identification of Invertebrate Pests at the Borders Using MinION Sequencing of DNA Barcodes. Genes, 12 (8). ISSN 2073-4425


Article Link: https://doi.org/10.3390/genes12081138


The rapid and accurate identification of invertebrate pests detected at the border is a challenging task. Current diagnostic methods used at the borders are mainly based on time consuming visual and microscopic examinations. Here, we demonstrate a rapid in-house workflow for DNA extraction, PCR amplification of the barcode region of the mitochondrial cytochrome oxidase subunit I (COI) gene and Oxford Nanopore Technologies (ONT) MinION sequencing of amplified products multiplexed after barcoding on ONT Flongle flow cells. A side-by-side comparison was conducted of DNA barcode sequencing-based identification and morphological identification of both large (>0.5 mm in length) and small (<0.5 mm in length) invertebrate specimens intercepted at the Australian border. DNA barcode sequencing results supported the morphological identification in most cases and enabled immature stages of invertebrates and their eggs to be identified more confidently. Results also showed that sequencing the COI barcode region using the ONT rapid sequencing principle is a cost-effective and field-adaptable approach for the rapid and accurate identification of invertebrate pests. Overall, the results suggest that MinION sequencing of DNA barcodes offers a complementary tool to the existing morphological diagnostic approaches and provides rapid, accurate, reliable and defendable evidence for identifying invertebrate pests at the border.

Item Type:Article
Business groups:Horticulture and Forestry Science
Keywords:cytochrome c oxidase subunit I MinION sequencing AgTech AgriTech invertebrate pests biosecurity
Subjects:Science > Entomology
Science > Invasive Species > Animals
Science > Zoology > Invertebrates > Insects
Science > Microbiology
Plant pests and diseases
Plant pests and diseases > Economic entomology
Technology > Technology (General)
Live Archive:05 Aug 2021 01:58
Last Modified:12 Sep 2022 07:03

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