{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,7,30]],"date-time":"2026-07-30T09:31:37Z","timestamp":1785403897086,"version":"3.56.0"},"reference-count":56,"publisher":"MDPI AG","issue":"8","license":[{"start":{"date-parts":[[2026,7,28]],"date-time":"2026-07-28T00:00:00Z","timestamp":1785196800000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"Science and Technology Innovation Project of Hebei Agriculture Research System","award":["HBCT2024150208"],"award-info":[{"award-number":["HBCT2024150208"]}]},{"name":"Hebei Academy of Agriculture and Forestry Sciences","award":["2026KJCXZX-SGS-9"],"award-info":[{"award-number":["2026KJCXZX-SGS-9"]}]},{"award":["2026KJCXZX-SGS-9"],"award-info":[{"award-number":["2026KJCXZX-SGS-9"]}],"id":[{"id":"https:\/\/ror.org\/051p3cy55","id-type":"ROR","asserted-by":"publisher"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Insects"],"abstract":"<jats:p>Leucoptera malifoliella (Lepidoptera: Lyonetiidae) is a quarantine pest of apple and other Rosaceae fruit trees whose range is expanding into new territories. Its minute size and morphological overlap with closely related Lyonetiidae make routine identification unreliable, especially for larvae and damaged specimens. We compared COI sequences from six common small Lepidoptera species found in orchards and designed the species-specific primer pair SXW-F\/SXW-R. The resulting polymerase chain reaction (PCR) assay amplifies an ~500 base pairs (bp) fragment exclusively from L. malifoliella; no product was detected in any of five non-target species. The reaction tolerates annealing temperatures of 50\u201358 \u00b0C and consistently detects the target across all life stages (first- to third-instar larvae, pupae, adults) and all adult tissues tested (antennae, head-thorax, abdomen, wings, legs). Detection sensitivity reaches 0.03 ng\/\u03bcL\u2014approximately one-thousandth of the DNA content of a single adult. This is the first species-specific COI (SS-COI) method reported for L. malifoliella. It furnishes a rapid, specific, and sensitive diagnostic tool for quarantine inspection, field monitoring, and integrated pest management (IPM) programs.<\/jats:p>","DOI":"10.3390\/insects17080778","type":"journal-article","created":{"date-parts":[[2026,7,28]],"date-time":"2026-07-28T11:45:26Z","timestamp":1785239126000},"page":"778","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":0,"title":["Species-Specific COI Primers for Rapid Molecular Identification of Leucoptera malifoliella"],"prefix":"10.3390","volume":"17","author":[{"ORCID":"https:\/\/orcid.org\/0000-0001-7411-8544","authenticated-orcid":false,"given":"Jiaqiang","family":"Zhao","sequence":"first","affiliation":[{"name":"Shijiazhuang Institute of Pomology, Hebei Academy of Agriculture and Forestry Sciences, Shijiazhuang 050061, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Qiang","family":"Xu","sequence":"additional","affiliation":[{"name":"Guangzhou Customs District Technology Center, Guangzhou 510623, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Guijie","family":"Chi","sequence":"additional","affiliation":[{"name":"Pingquan Municipal Bureau of Agriculture and Rural Affairs, Chengde 067500, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Shengping","family":"Zhang","sequence":"additional","affiliation":[{"name":"Shijiazhuang Institute of Pomology, Hebei Academy of Agriculture and Forestry Sciences, Shijiazhuang 050061, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Ruitao","family":"Yu","sequence":"additional","affiliation":[{"name":"Shijiazhuang Institute of Pomology, Hebei Academy of Agriculture and Forestry Sciences, Shijiazhuang 050061, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0009-0005-7788-9164","authenticated-orcid":false,"given":"Shihang","family":"Zhao","sequence":"additional","affiliation":[{"name":"Shijiazhuang Institute of Pomology, Hebei Academy of Agriculture and Forestry Sciences, Shijiazhuang 050061, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Qi","family":"Gao","sequence":"additional","affiliation":[{"name":"Shijiazhuang Institute of Pomology, Hebei Academy of Agriculture and Forestry Sciences, Shijiazhuang 050061, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Zhaohui","family":"Yang","sequence":"additional","affiliation":[{"name":"Hebei Provincial Center of Agro-Product Safety and Quality, Shijiazhuang 050061, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0009-0004-0950-8529","authenticated-orcid":false,"given":"Guoliang","family":"Xu","sequence":"additional","affiliation":[{"name":"Shijiazhuang Institute of Pomology, Hebei Academy of Agriculture and Forestry Sciences, Shijiazhuang 050061, China"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2026,7,28]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"53","DOI":"10.11646\/zootaxa.1245.1.2","article-title":"Lyonetiidae of Turkey with notes on their distribution and zoogeography (Lepidoptera)","volume":"1245","author":"Seven","year":"2006","journal-title":"Zootaxa"},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"11","DOI":"10.30574\/gscbps.2017.1.1.0008","article-title":"Quantitative analysis of the causes of outbreaks in leaf-mining insects: Application to the common leaf-miner Leucoptera malifoliella (Lepidoptera: Lyonetiidae)","volume":"1","year":"2017","journal-title":"GSC Biol. 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