{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,8,13]],"date-time":"2026-08-13T09:27:05Z","timestamp":1786613225971,"version":"build-2736575974"},"reference-count":31,"publisher":"MDPI AG","issue":"8","license":[{"start":{"date-parts":[[2026,8,13]],"date-time":"2026-08-13T00:00:00Z","timestamp":1786579200000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"Key Research and Development Program of Xinjiang Uygur Autonomous Region, China","award":["2026B02001"],"award-info":[{"award-number":["2026B02001"]}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Insects"],"abstract":"<jats:p>Ips hauseri Reitter is the most important wood-boring pest of natural Tianshan spruce (Picea schrenkiana) forests and is listed as an A2 quarantine pest by the European and Mediterranean Plant Protection Organization (EPPO). Because the beetle is small and cryptic and its immature stages cannot be reliably identified by morphology, a rapid molecular identification method is urgently needed. Here, we targeted the mitochondrial cytochrome c oxidase subunit I (COI) gene, designed a set of I. hauseri-specific LAMP primers, and established a visually interpretable LAMP assay using a WarmStart colorimetric system. The optimal reaction conditions were 63.8 \u00b0C for 35 min. The assay amplified only I. hauseri, with no cross-reaction with the related species Pityogenes spessivtsevi, Ips typographus, Ips subelongatus, and Hylurgus ligniperda; its limit of detection was 100 pg of genomic DNA; and it amplified efficiently from adults, larvae, pupae, and a single egg. Positive results could be scored directly by eye from the red-to-yellow colour change in the reaction, requiring no precision instruments at any step. Using a non-destructive alkaline crude-DNA extraction (HotSHOT), the assay also amplified I. hauseri directly from single adults and larvae, confirming its compatibility with a rapid, field-simple sample preparation. The LAMP assay was rapid, specific within the tested panel, and simple. However, broader validation with additional sympatric non-target species and geographically distinct populations of I. hauseri is needed before routine field or quarantine application.<\/jats:p>","DOI":"10.3390\/insects17080838","type":"journal-article","created":{"date-parts":[[2026,8,13]],"date-time":"2026-08-13T08:06:16Z","timestamp":1786608376000},"page":"838","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":0,"title":["Development of a Loop-Mediated Isothermal Amplification (LAMP) Assay for Rapid Molecular Identification of the Bark Beetle Ips hauseri (Coleoptera: Curculionidae)"],"prefix":"10.3390","volume":"17","author":[{"given":"Jipeng","family":"Jiao","sequence":"first","affiliation":[{"name":"College of Forestry and Landscape Architecture, Xinjiang Agricultural University, Urumqi 830052, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Wei","family":"Zhang","sequence":"additional","affiliation":[{"name":"College of Forestry and Landscape Architecture, Xinjiang Agricultural University, Urumqi 830052, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Liang","family":"Yan","sequence":"additional","affiliation":[{"name":"Midong Branch, Eastern Tianshan State-owned Forest Administration Bureau, Urumqi 831400, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yang","family":"Zhou","sequence":"additional","affiliation":[{"name":"College of Forestry and Landscape Architecture, Xinjiang Agricultural University, Urumqi 830052, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Shucheng","family":"Li","sequence":"additional","affiliation":[{"name":"College of Forestry and Landscape Architecture, Xinjiang Agricultural University, Urumqi 830052, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-0333-0681","authenticated-orcid":false,"given":"Lili","family":"Ren","sequence":"additional","affiliation":[{"name":"Beijing Key Laboratory for Forest Pest Control, Beijing Forestry University, Beijing 100083, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Adil","family":"Sattar","sequence":"additional","affiliation":[{"name":"College of Forestry and Landscape Architecture, Xinjiang Agricultural University, Urumqi 830052, China"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2026,8,13]]},"reference":[{"key":"ref_1","first-page":"9","article-title":"Distribution patterns of Picea schrenkiana var. tianschanica population in Tianshan Mountains","volume":"22","author":"Liu","year":"2011","journal-title":"Chin. J. Appl. Ecol."},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"267","DOI":"10.1016\/j.foreco.2004.02.063","article-title":"Age structure of Picea schrenkiana forest along an altitudinal gradient in the central Tianshan Mountains, northwestern China","volume":"196","author":"Wang","year":"2004","journal-title":"For. Ecol. Manag."},{"key":"ref_3","doi-asserted-by":"crossref","unstructured":"EPPO (2005). Data sheets on quarantine pests: Ips hauseri. EPPO Bull., 35, 450\u2013452.","DOI":"10.1111\/j.1365-2338.2005.00881.x"},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"257","DOI":"10.1007\/s11157-024-09692-5","article-title":"Understanding bark beetle outbreaks: Exploring the impact of changing temperature regimes, droughts, forest structure, and prospects for future forest pest management","volume":"23","author":"Singh","year":"2024","journal-title":"Rev. Environ. Sci. Biotechnol."},{"key":"ref_5","unstructured":"Yin, H.F., Huang, F.S., and Li, Z.L. (1984). Economic Insect Fauna of China, Fasc. 29, Coleoptera: Scolytidae, Science Press."},{"key":"ref_6","doi-asserted-by":"crossref","unstructured":"Albo, J.E., Marelli, J.P., and Puig, A.S. (2019). Rapid molecular identification of Scolytinae (Coleoptera: Curculionidae). Int. J. Mol. Sci., 20.","DOI":"10.3390\/ijms20235944"},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"313","DOI":"10.1098\/rspb.2002.2218","article-title":"Biological identifications through DNA barcodes","volume":"270","author":"Hebert","year":"2003","journal-title":"Proc. R. Soc. B Biol. Sci."},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"1473","DOI":"10.1007\/s41348-024-00902-4","article-title":"Rapid identification of Ips sexdentatus (Boerner, 1766) (Curculionidae) from adults and frass with real-time PCR based on probe technology","volume":"131","author":"Rizzo","year":"2024","journal-title":"J. Plant Dis. Prot."},{"key":"ref_9","doi-asserted-by":"crossref","unstructured":"Vulchi, R., Daane, K.M., and Wenger, J.A. (2021). Development of DNA melt curve analysis for the identification of lepidopteran pests in almonds and pistachios. Insects, 12.","DOI":"10.3390\/insects12060553"},{"key":"ref_10","doi-asserted-by":"crossref","unstructured":"Nagpal, K., Jasrotia, P., Jaglan, M.S., Kashyap, P.L., Maanju, S., and Kumar, S. (2025). Molecular detection tool for prompt, reliable and precise diagnosis of the rice weevil, Sitophilus oryzae (Linnaeus) infestation in wheat. Front. Plant Sci., 16.","DOI":"10.3389\/fpls.2025.1628692"},{"key":"ref_11","doi-asserted-by":"crossref","unstructured":"Jangra, S., Ghosh, A., Mukherjee, S., Baranwal, V.K., and Dietzgen, R.G. (2022). Development of a polymerase spiral reaction-based isothermal assay for rapid identification of Thrips palmi. Front. Mol. Biosci., 9.","DOI":"10.3389\/fmolb.2022.853339"},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"e63","DOI":"10.1093\/nar\/28.12.e63","article-title":"Loop-mediated isothermal amplification of DNA","volume":"28","author":"Notomi","year":"2000","journal-title":"Nucleic Acids Res."},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"877","DOI":"10.1038\/nprot.2008.57","article-title":"Loop-mediated isothermal amplification (LAMP) of gene sequences and simple visual detection of products","volume":"3","author":"Tomita","year":"2008","journal-title":"Nat. Protoc."},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"21229","DOI":"10.1038\/s41598-020-77928-9","article-title":"A diagnostic LAMP assay for the destructive grapevine insect pest, phylloxera (Daktulosphaira vitifoliae)","volume":"10","author":"Agarwal","year":"2020","journal-title":"Sci. Rep."},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"1116","DOI":"10.1038\/s41598-021-04496-x","article-title":"A diagnostic LAMP assay for rapid identification of an invasive plant pest, fall armyworm Spodoptera frugiperda (Lepidoptera: Noctuidae)","volume":"12","author":"Agarwal","year":"2022","journal-title":"Sci. Rep."},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"100029","DOI":"10.1016\/j.cris.2022.100029","article-title":"A loop-mediated isothermal amplification (LAMP) assay for rapid identification of Ceratitis capitata and related species","volume":"2","author":"Dermauw","year":"2022","journal-title":"Curr. Res. Insect Sci."},{"key":"ref_17","doi-asserted-by":"crossref","unstructured":"Nam, H.Y., Kim, J.H., Lee, S.H., Heckel, D.G., and Kim, J. (2021). Development of a LAMP-based molecular species diagnosis method for four major agricultural pests in the genus Spodoptera (Lepidoptera: Noctuidae). Insects, 12.","DOI":"10.3390\/insects12100883"},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"2911","DOI":"10.1093\/jee\/toaa206","article-title":"Molecular identification of Anoplophora glabripennis (Coleoptera: Cerambycidae) from frass by loop-mediated isothermal amplification","volume":"113","author":"Rizzo","year":"2020","journal-title":"J. Econ. Entomol."},{"key":"ref_19","doi-asserted-by":"crossref","unstructured":"Rizzo, D., Moricca, S., Bracalini, M., Benigno, A., Bernardo, U., Luchi, N., Da Lio, D., Nugnes, F., Cappellini, G., and Salemi, C. (2021). Rapid detection of Pityophthorus juglandis (Blackman) (Coleoptera, Curculionidae) with the loop-mediated isothermal amplification (LAMP) method. Plants, 10.","DOI":"10.3390\/plants10061048"},{"key":"ref_20","doi-asserted-by":"crossref","unstructured":"Jiao, J., Ren, L., Chen, R., Tao, J., and Luo, Y. (2022). A LAMP assay for the detection of Thecodiplosis japonensis, an alien gall midge species pest of pine trees. Insects, 13.","DOI":"10.3390\/insects13060540"},{"key":"ref_21","unstructured":"Zhang, S.M. (2009). The Molecular Phylogenetic Research of Ips typographus Based on Partial Sequences of mtDNA-12S rRNA, COI, and COII Genes. [Master\u2019s Thesis, Northeast Forestry University]."},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"545","DOI":"10.1146\/annurev.ecolsys.37.091305.110018","article-title":"Incorporating molecular evolution into phylogenetic analysis, and a new compilation of conserved polymerase chain reaction primers for animal mitochondrial DNA","volume":"37","author":"Simon","year":"2006","journal-title":"Annu. Rev. Ecol. Evol. Syst."},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"3022","DOI":"10.1093\/molbev\/msab120","article-title":"MEGA11: Molecular Evolutionary Genetics Analysis version 11","volume":"38","author":"Tamura","year":"2021","journal-title":"Mol. Biol. Evol."},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"52","DOI":"10.2144\/00291bm09","article-title":"Preparation of PCR-quality mouse genomic DNA with hot sodium hydroxide and tris (HotSHOT)","volume":"29","author":"Truett","year":"2000","journal-title":"BioTechniques"},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"198","DOI":"10.1093\/mollus\/eyy011","article-title":"Development and evaluation of hotshot protocols for cost- and time-effective extraction of PCR-ready DNA from single freshwater mussel larvae (Bivalvia: Unionida)","volume":"84","author":"Zieritz","year":"2018","journal-title":"J. Molluscan Stud."},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"287","DOI":"10.1139\/gen-2018-0045","article-title":"Non-destructive DNA extraction methods for entomophagous insects with emphasis on biological control","volume":"62","author":"Gallou","year":"2019","journal-title":"Genome"},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"59","DOI":"10.2144\/000114253","article-title":"Visual detection of isothermal nucleic acid amplification using pH-sensitive dyes","volume":"58","author":"Tanner","year":"2015","journal-title":"BioTechniques"},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"567","DOI":"10.1094\/PHYTO-08-22-0319-R","article-title":"Field-portable, rapid, and low-cost RT-LAMP assay for the detection of tomato chlorotic spot virus","volume":"113","author":"Yilmaz","year":"2023","journal-title":"Phytopathology"},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"626","DOI":"10.1111\/jam.13647","article-title":"Loop-mediated isothermal amplification (LAMP): A versatile technique for detection of micro-organisms","volume":"124","author":"Wong","year":"2018","journal-title":"J. Appl. Microbiol."},{"key":"ref_30","doi-asserted-by":"crossref","unstructured":"Huang, B., Montgomery, B.L., Adamczyk, R., Ehlers, G., van den Hurk, A.F., and Warrilow, D. (2020). A LAMP-based colorimetric assay to expedite field surveillance of the invasive mosquito species Aedes aegypti and Aedes albopictus. PLoS Negl. Trop. Dis., 14.","DOI":"10.1371\/journal.pntd.0008130"},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"223","DOI":"10.1006\/mcpr.2002.0415","article-title":"Accelerated reaction by loop-mediated isothermal amplification using loop primers","volume":"16","author":"Nagamine","year":"2002","journal-title":"Mol. Cell. Probes"}],"container-title":["Insects"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/2075-4450\/17\/8\/838\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2026,8,13]],"date-time":"2026-08-13T08:32:29Z","timestamp":1786609949000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/2075-4450\/17\/8\/838"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2026,8,13]]},"references-count":31,"journal-issue":{"issue":"8","published-online":{"date-parts":[[2026,8]]}},"alternative-id":["insects17080838"],"URL":"https:\/\/doi.org\/10.3390\/insects17080838","relation":{},"ISSN":["2075-4450"],"issn-type":[{"value":"2075-4450","type":"electronic"}],"subject":[],"published":{"date-parts":[[2026,8,13]]}}}