{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,21]],"date-time":"2026-10-21T17:54:19Z","timestamp":1792605259660,"version":"4.5.0"},"reference-count":51,"publisher":"Wiley","issue":"12","license":[{"start":{"date-parts":[[2023,9,7]],"date-time":"2023-09-07T00:00:00Z","timestamp":1694044800000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/onlinelibrary.wiley.com\/termsAndConditions#vor"}],"funder":[{"DOI":"10.13039\/501100012166","name":"National Key Research and Development Program of China","doi-asserted-by":"publisher","award":["2022YFC2601500"],"award-info":[{"award-number":["2022YFC2601500"]}],"id":[{"id":"10.13039\/501100012166","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":["scijournals.onlinelibrary.wiley.com"],"crossmark-restriction":true},"short-container-title":["Pest Management Science"],"published-print":{"date-parts":[[2023,12]]},"abstract":"<jats:title>Abstract<\/jats:title>\n                  <jats:sec>\n                    <jats:title>BACKGROUND<\/jats:title>\n                    <jats:p>\n                      Khapra beetle (\n                      <jats:italic>Trogoderma granarium<\/jats:italic>\n                      Everts), one of the most important quarantine pests globally, is capable of causing severe infestation and huge economic loss to stored grain, and its interception rate has increased in major global trade countries over the past few years. However, difficulties remain in distinguishing this species with similar ones. In order to assist border ports and warehouses in khapra beetle's effective rapid identification as well as pest control at the early stages of monitoring or interception, we herein developed a new and rapid visual detection assay for\n                      <jats:italic>T.\u2009granarium<\/jats:italic>\n                      based on recombinase polymerase amplification (RPA) and the CRISPR\/Cas12a system.\n                    <\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>RESULTS<\/jats:title>\n                    <jats:p>\n                      We designed and selected the first khapra beetle\u2010specific RPA primers and crRNA, and optimized the visualization reaction system (Cas12a\/CrRNA\u2009=\u2009100\u2009n\n                      <jats:sc>M<\/jats:sc>\n                      \/500\u2009n\n                      <jats:sc>M<\/jats:sc>\n                      ). With only a 37\u2009\u00b0C\u2010heat\u2010source and a blue light torch, RPA and CRISPR\/CAS12a\u2010based visualization assays can be completed within 40\u2009min to differentiate between khapra beetle and nine similar Dermestidae species. After DNA extraction using a kit (4\u20135\u2009h) or a simple method (5\u2009min), the specific amplicons were obtained after a 15\u2009min RPA reaction at 37\u2009\u00b0C, followed by a 15\u2009min color reaction under 37\u2009\u00b0C in dark conditions using a CRISPR\/CAS12a system and a fluorescent probe (5\u2032\u2010FAM\/3\u2032\u2010BHQ1 labeled). This method is ingenious to low levels of DNA (10\n                      <jats:sup>\u22121<\/jats:sup>\n                      \u2009ng\u2009\u03bcL\n                      <jats:sup>\u22121<\/jats:sup>\n                      ) and meets the sensitivity requirements for detecting a single khapra beetle's egg (\u22480.7\u2009mm).\n                    <\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>CONCLUSION<\/jats:title>\n                    <jats:p>Our specificity and sensitivity analysis inferred that the present visualization system is effective to quickly and uniquely detect khapra beetle at room temperature (37\u2009\u00b0C), thereby preventing this species before they spread widely. Our study is suitable for being pushed forward in storage pest management, and provides value as a reference for monitoring and identification of other pests. \u00a9 2023 Society of Chemical Industry.<\/jats:p>\n                  <\/jats:sec>","DOI":"10.1002\/ps.7739","type":"journal-article","created":{"date-parts":[[2023,8,22]],"date-time":"2023-08-22T05:16:47Z","timestamp":1692681407000},"page":"5304-5311","update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":14,"title":["New and rapid visual detection assay for\n                    <i>Trogoderma granarium<\/i>\n                    everts based on recombinase polymerase amplification and\n                    <scp>CRISPR<\/scp>\n                    \/Cas12a"],"prefix":"10.1002","volume":"79","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-5422-1314","authenticated-orcid":false,"given":"Lingyu","family":"Zeng","sequence":"first","affiliation":[{"name":"Department of Plant Biosecurity, College of Plant Protection China Agricultural University  Beijing P. R. China"},{"name":"Key Laboratory of Surveillance and Management for Plant Quarantine Pests Ministry of Agriculture and Rural Affairs  Beijing P. R. China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Sizhu","family":"Zheng","sequence":"additional","affiliation":[{"name":"Suzhou Customs District  Suzhou P. R. China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Vaclav","family":"Stejskal","sequence":"additional","affiliation":[{"name":"Crop Research Institute, Drnovsk\u00e1, Prague  Czech Republic"},{"name":"Faculty of Agrobiology, Food and Natural Resources Czech University of Life Sciences  Prague Czech Republic"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"George","family":"Opit","sequence":"additional","affiliation":[{"name":"Department of Entomology and Plant Pathology Oklahoma State University, 127 Noble Research Center  Stillwater USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Radek","family":"Aulicky","sequence":"additional","affiliation":[{"name":"Crop Research Institute, Drnovsk\u00e1, Prague  Czech Republic"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-1281-2108","authenticated-orcid":false,"given":"Zhihong","family":"Li","sequence":"additional","affiliation":[{"name":"Department of Plant Biosecurity, College of Plant Protection China Agricultural University  Beijing P. R. China"},{"name":"Key Laboratory of Surveillance and Management for Plant Quarantine Pests Ministry of Agriculture and Rural Affairs  Beijing P. R. China"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"311","published-online":{"date-parts":[[2023,9,7]]},"reference":[{"key":"e_1_2_9_2_1","volume-title":"Diagnostic Protocols for Regulated Pests, DP 3: Trogoderma Granarium Everts","author":"IPPC ISPM 27","year":"2016"},{"key":"e_1_2_9_3_1","first-page":"48","article-title":"Quarantine importance of Trogoderma pests","volume":"23","author":"Liu HF","year":"2009","journal-title":"Plant Quar"},{"key":"e_1_2_9_4_1","doi-asserted-by":"publisher","DOI":"10.1111\/epp.12080"},{"key":"e_1_2_9_5_1","doi-asserted-by":"publisher","DOI":"10.1146\/annurev-ento-011118-111804"},{"key":"e_1_2_9_6_1","unstructured":"AthanassiouCG Trogoderma granarium(khapra beetle).https:\/\/www.cabidigitallibrary.org\/doi\/epdf\/10.1079\/cabicompendium.55010[10 March 2023]."},{"key":"e_1_2_9_7_1","doi-asserted-by":"publisher","DOI":"10.1016\/0308-8146(92)90189-9"},{"key":"e_1_2_9_8_1","doi-asserted-by":"publisher","DOI":"10.1002\/ps.6591"},{"key":"e_1_2_9_9_1","doi-asserted-by":"publisher","DOI":"10.3390\/insects12050442"},{"key":"e_1_2_9_10_1","volume-title":"Illustrated Identification Keys for Trogoderma granarium, T. glabrum, T. inclusum and T. 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