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LAMP (Loop-mediated isothermal amplification) assay for rapid identification of Varroa mites
Varroa mites are serious pests of European honeybees (Apis mellifera). For detection of Varroa mite, a new molecular LAMP-based assay has been developed, which retains the body of the mite intact for morphological identification. Six novel Varroa LAMP primers were designed from existing DNA sequence...
Autores principales: | , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
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Nature Publishing Group UK
2023
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10366197/ https://www.ncbi.nlm.nih.gov/pubmed/37488147 http://dx.doi.org/10.1038/s41598-023-38860-w |
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author | Rako, Lea Agarwal, Arati Eow, Lixin Roberts, John M. K. Rodoni, Brendan C. Blacket, Mark J. |
author_facet | Rako, Lea Agarwal, Arati Eow, Lixin Roberts, John M. K. Rodoni, Brendan C. Blacket, Mark J. |
author_sort | Rako, Lea |
collection | PubMed |
description | Varroa mites are serious pests of European honeybees (Apis mellifera). For detection of Varroa mite, a new molecular LAMP-based assay has been developed, which retains the body of the mite intact for morphological identification. Six novel Varroa LAMP primers were designed from existing DNA sequences of the COI locus to target V. destructor and V. jacobsoni, providing the ability to tell them apart from other non-target beehive associated mite and insect species. This LAMP assay is specific in detecting these Varroa species and has been tested on specimens originating from multiple countries. It produces amplification of V. destructor and V. jacobsoni in 16 ± 3.4 min with an anneal derivative of 78 ± 0.5 °C whilst another Varroa species,V. underwoodi, showed late amplification. A gBlock gene fragment, used here as a positive control has a different anneal derivative of 80 °C. Three non-destructive DNA extraction methods (HotShot, QuickExtract and Xtract) were tested and found to be suitable for use in the field. The LAMP assay was sensitive to very low levels of Varroa DNA, down to 0.24 picogram (~ 1 × 10 copies/µL of Varroa gBlock). This is a new molecular tool for rapid and accurate detection and identification of Varroa mites for pest management, in areas where these mites do not occur. |
format | Online Article Text |
id | pubmed-10366197 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2023 |
publisher | Nature Publishing Group UK |
record_format | MEDLINE/PubMed |
spelling | pubmed-103661972023-07-26 LAMP (Loop-mediated isothermal amplification) assay for rapid identification of Varroa mites Rako, Lea Agarwal, Arati Eow, Lixin Roberts, John M. K. Rodoni, Brendan C. Blacket, Mark J. Sci Rep Article Varroa mites are serious pests of European honeybees (Apis mellifera). For detection of Varroa mite, a new molecular LAMP-based assay has been developed, which retains the body of the mite intact for morphological identification. Six novel Varroa LAMP primers were designed from existing DNA sequences of the COI locus to target V. destructor and V. jacobsoni, providing the ability to tell them apart from other non-target beehive associated mite and insect species. This LAMP assay is specific in detecting these Varroa species and has been tested on specimens originating from multiple countries. It produces amplification of V. destructor and V. jacobsoni in 16 ± 3.4 min with an anneal derivative of 78 ± 0.5 °C whilst another Varroa species,V. underwoodi, showed late amplification. A gBlock gene fragment, used here as a positive control has a different anneal derivative of 80 °C. Three non-destructive DNA extraction methods (HotShot, QuickExtract and Xtract) were tested and found to be suitable for use in the field. The LAMP assay was sensitive to very low levels of Varroa DNA, down to 0.24 picogram (~ 1 × 10 copies/µL of Varroa gBlock). This is a new molecular tool for rapid and accurate detection and identification of Varroa mites for pest management, in areas where these mites do not occur. Nature Publishing Group UK 2023-07-24 /pmc/articles/PMC10366197/ /pubmed/37488147 http://dx.doi.org/10.1038/s41598-023-38860-w Text en © Crown 2023 https://creativecommons.org/licenses/by/4.0/Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by/4.0/ (https://creativecommons.org/licenses/by/4.0/) . |
spellingShingle | Article Rako, Lea Agarwal, Arati Eow, Lixin Roberts, John M. K. Rodoni, Brendan C. Blacket, Mark J. LAMP (Loop-mediated isothermal amplification) assay for rapid identification of Varroa mites |
title | LAMP (Loop-mediated isothermal amplification) assay for rapid identification of Varroa mites |
title_full | LAMP (Loop-mediated isothermal amplification) assay for rapid identification of Varroa mites |
title_fullStr | LAMP (Loop-mediated isothermal amplification) assay for rapid identification of Varroa mites |
title_full_unstemmed | LAMP (Loop-mediated isothermal amplification) assay for rapid identification of Varroa mites |
title_short | LAMP (Loop-mediated isothermal amplification) assay for rapid identification of Varroa mites |
title_sort | lamp (loop-mediated isothermal amplification) assay for rapid identification of varroa mites |
topic | Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10366197/ https://www.ncbi.nlm.nih.gov/pubmed/37488147 http://dx.doi.org/10.1038/s41598-023-38860-w |
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