{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,24]],"date-time":"2026-10-24T01:29:16Z","timestamp":1792805356138,"version":"4.5.0"},"reference-count":32,"publisher":"MDPI AG","issue":"18","license":[{"start":{"date-parts":[[2023,9,15]],"date-time":"2023-09-15T00:00:00Z","timestamp":1694736000000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/501100012166","name":"National Key Research and Development Program of China","doi-asserted-by":"publisher","award":["2022YFD1400100"],"award-info":[{"award-number":["2022YFD1400100"]}],"id":[{"id":"10.13039\/501100012166","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100012166","name":"National Key Research and Development Program of China","doi-asserted-by":"publisher","award":["2022YFE0139500"],"award-info":[{"award-number":["2022YFE0139500"]}],"id":[{"id":"10.13039\/501100012166","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100012166","name":"National Key Research and Development Program of China","doi-asserted-by":"publisher","award":["32072377"],"award-info":[{"award-number":["32072377"]}],"id":[{"id":"10.13039\/501100012166","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100012166","name":"National Key Research and Development Program of China","doi-asserted-by":"publisher","award":["32102185"],"award-info":[{"award-number":["32102185"]}],"id":[{"id":"10.13039\/501100012166","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100012166","name":"National Key Research and Development Program of China","doi-asserted-by":"publisher","award":["CAAS-ASTIP-G2022-IFST-01"],"award-info":[{"award-number":["CAAS-ASTIP-G2022-IFST-01"]}],"id":[{"id":"10.13039\/501100012166","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100012166","name":"National Key Research and Development Program of China","doi-asserted-by":"publisher","award":["Y2022GH15"],"award-info":[{"award-number":["Y2022GH15"]}],"id":[{"id":"10.13039\/501100012166","id-type":"DOI","asserted-by":"publisher"}]},{"name":"National Natural Science Foundation of China","award":["2022YFD1400100"],"award-info":[{"award-number":["2022YFD1400100"]}],"id":[{"id":"https:\/\/ror.org\/01h0zpd94","id-type":"ROR","asserted-by":"crossref"}]},{"name":"National Natural Science Foundation of China","award":["2022YFE0139500"],"award-info":[{"award-number":["2022YFE0139500"]}],"id":[{"id":"https:\/\/ror.org\/01h0zpd94","id-type":"ROR","asserted-by":"crossref"}]},{"name":"National Natural Science Foundation of China","award":["32072377"],"award-info":[{"award-number":["32072377"]}],"id":[{"id":"https:\/\/ror.org\/01h0zpd94","id-type":"ROR","asserted-by":"crossref"}]},{"name":"National Natural Science Foundation of China","award":["32102185"],"award-info":[{"award-number":["32102185"]}],"id":[{"id":"https:\/\/ror.org\/01h0zpd94","id-type":"ROR","asserted-by":"crossref"}]},{"name":"National Natural Science Foundation of China","award":["CAAS-ASTIP-G2022-IFST-01"],"award-info":[{"award-number":["CAAS-ASTIP-G2022-IFST-01"]}],"id":[{"id":"https:\/\/ror.org\/01h0zpd94","id-type":"ROR","asserted-by":"crossref"}]},{"name":"National Natural Science Foundation of China","award":["Y2022GH15"],"award-info":[{"award-number":["Y2022GH15"]}],"id":[{"id":"https:\/\/ror.org\/01h0zpd94","id-type":"ROR","asserted-by":"crossref"}]},{"name":"Agricultural Science and Technology Innovation Program of Institute of Food Science and Technology","award":["2022YFD1400100"],"award-info":[{"award-number":["2022YFD1400100"]}]},{"name":"Agricultural Science and Technology Innovation Program of Institute of Food Science and Technology","award":["2022YFE0139500"],"award-info":[{"award-number":["2022YFE0139500"]}]},{"name":"Agricultural Science and Technology Innovation Program of Institute of Food Science and Technology","award":["32072377"],"award-info":[{"award-number":["32072377"]}]},{"name":"Agricultural Science and Technology Innovation Program of Institute of Food Science and Technology","award":["32102185"],"award-info":[{"award-number":["32102185"]}]},{"name":"Agricultural Science and Technology Innovation Program of Institute of Food Science and Technology","award":["CAAS-ASTIP-G2022-IFST-01"],"award-info":[{"award-number":["CAAS-ASTIP-G2022-IFST-01"]}]},{"name":"Agricultural Science and Technology Innovation Program of Institute of Food Science and Technology","award":["Y2022GH15"],"award-info":[{"award-number":["Y2022GH15"]}]},{"name":"Central Public-interest Scientific Institution Basal Research Fund","award":["2022YFD1400100"],"award-info":[{"award-number":["2022YFD1400100"]}]},{"name":"Central Public-interest Scientific Institution Basal Research Fund","award":["2022YFE0139500"],"award-info":[{"award-number":["2022YFE0139500"]}]},{"name":"Central Public-interest Scientific Institution Basal Research Fund","award":["32072377"],"award-info":[{"award-number":["32072377"]}]},{"name":"Central Public-interest Scientific Institution Basal Research Fund","award":["32102185"],"award-info":[{"award-number":["32102185"]}]},{"name":"Central Public-interest Scientific Institution Basal Research Fund","award":["CAAS-ASTIP-G2022-IFST-01"],"award-info":[{"award-number":["CAAS-ASTIP-G2022-IFST-01"]}]},{"name":"Central Public-interest Scientific Institution Basal Research Fund","award":["Y2022GH15"],"award-info":[{"award-number":["Y2022GH15"]}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["IJMS"],"abstract":"<jats:p>Fusarium head blight (FHB) is a global cereal disease caused by a complex of Fusarium species. Both Fusarium graminearum and F. asiaticum are the causal agents of FHB in China. F. asiaticum is the predominant species in the Middle\u2013Lower Reaches of the Yangtze River (MLRYR) and southwest China. Therefore, detecting F. asiaticum in a timely manner is crucial for controlling the disease and preventing mycotoxins from entering the food chain. Here, we combined rapid genomic DNA extraction, recombinase polymerase amplification, Cas12a cleavage, and lateral flow detection techniques to develop a method for the rapid detection of F. asiaticum. The reaction conditions were optimized to provide a rapid, sensitive, and cost-effective method for F. asiaticum detection. The optimized method demonstrated exceptional specificity in detecting F. asiaticum while not detecting any of the 14 other Fusarium strains and 3 non-Fusarium species. Additionally, it could detect F. asiaticum DNA at concentrations as low as 20 ag\/\u03bcL, allowing for the diagnosis of F. asiaticum infection in maize and wheat kernels even after 3 days of inoculation. The developed assay will provide an efficient and robust detection platform to accelerate plant pathogen detection.<\/jats:p>","DOI":"10.3390\/ijms241814134","type":"journal-article","created":{"date-parts":[[2023,9,15]],"date-time":"2023-09-15T02:54:13Z","timestamp":1694746453000},"page":"14134","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":14,"title":["Rapid and Sensitive Detection of Toxigenic Fusarium asiaticum Integrating Recombinase Polymerase Amplification, CRISPR\/Cas12a, and Lateral Flow Techniques"],"prefix":"10.3390","volume":"24","author":[{"given":"Jun","family":"Zhang","sequence":"first","affiliation":[{"name":"State Key Laboratory for Biology of Plant Diseases and Insect Pests, Institute of Plant Protection, Chinese Academy of Agricultural Sciences, Beijing 100193, China"},{"name":"Institute of Food Science and Technology, Chinese Academy of Agricultural Sciences, Key Laboratory of Agro-Products Quality and Safety Control in Storage and Transport Process, Ministry of Agriculture and Rural Affairs, Beijing 100193, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Xiaoyan","family":"Liang","sequence":"additional","affiliation":[{"name":"Institute of Food Science and Technology, Chinese Academy of Agricultural Sciences, Key Laboratory of Agro-Products Quality and Safety Control in Storage and Transport Process, Ministry of Agriculture and Rural Affairs, Beijing 100193, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Hao","family":"Zhang","sequence":"additional","affiliation":[{"name":"State Key Laboratory for Biology of Plant Diseases and Insect Pests, Institute of Plant Protection, Chinese Academy of Agricultural Sciences, Beijing 100193, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Shumila","family":"Ishfaq","sequence":"additional","affiliation":[{"name":"Institute of Food Science and Technology, Chinese Academy of Agricultural Sciences, Key Laboratory of Agro-Products Quality and Safety Control in Storage and Transport Process, Ministry of Agriculture and Rural Affairs, Beijing 100193, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Kaifei","family":"Xi","sequence":"additional","affiliation":[{"name":"Institute of Food Science and Technology, Chinese Academy of Agricultural Sciences, Key Laboratory of Agro-Products Quality and Safety Control in Storage and Transport Process, Ministry of Agriculture and Rural Affairs, Beijing 100193, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-5311-7331","authenticated-orcid":false,"given":"Xueping","family":"Zhou","sequence":"additional","affiliation":[{"name":"State Key Laboratory for Biology of Plant Diseases and Insect Pests, Institute of Plant Protection, Chinese Academy of Agricultural Sciences, Beijing 100193, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Xiuling","family":"Yang","sequence":"additional","affiliation":[{"name":"State Key Laboratory for Biology of Plant Diseases and Insect Pests, Institute of Plant Protection, Chinese Academy of Agricultural Sciences, Beijing 100193, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-6411-6227","authenticated-orcid":false,"given":"Wei","family":"Guo","sequence":"additional","affiliation":[{"name":"Institute of Food Science and Technology, Chinese Academy of Agricultural Sciences, Key Laboratory of Agro-Products Quality and Safety Control in Storage and Transport Process, Ministry of Agriculture and Rural Affairs, Beijing 100193, China"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2023,9,15]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"515","DOI":"10.1111\/j.1364-3703.2004.00252.x","article-title":"Heading for disaster: Fusarium graminearum on cereal crops","volume":"5","author":"Goswami","year":"2004","journal-title":"Mol. Plant Pathol."},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"1096","DOI":"10.1016\/j.fgb.2011.09.002","article-title":"Novel Fusarium head blight pathogens from Nepal and Louisiana revealed by multilocus genealogical concordance","volume":"48","author":"Sarver","year":"2011","journal-title":"Fungal Genet. Biol."},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"100","DOI":"10.1016\/j.cropro.2015.02.025","article-title":"Management of Fusarium head blight of wheat and barley","volume":"73","author":"Wegulo","year":"2015","journal-title":"Crop Prot."},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"453","DOI":"10.1080\/19440049.2014.984244","article-title":"Biogeography of Fusarium graminearum species complex and chemotypes: A review","volume":"32","author":"Zhang","year":"2015","journal-title":"Food Addit. Contam. Part A Chem. Anal. Control Expo. Risk Assess."},{"key":"ref_5","doi-asserted-by":"crossref","unstructured":"Zhang, H., Van der Lee, T., Waalwijk, C., Chen, W., Xu, J., Xu, J., Zhang, Y., and Feng, J. (2012). Population analysis of the Fusarium graminearum species complex from wheat in China show a shift to more aggressive isolates. PLoS ONE, 7.","DOI":"10.1371\/journal.pone.0031722"},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"2830","DOI":"10.1094\/PDIS-01-21-0013-RE","article-title":"The distribution of Fusarium graminearum and Fusarium asiaticum causing Fusarium Head Blight of wheat in relation to climate and cropping system","volume":"105","author":"Xu","year":"2021","journal-title":"Plant Dis."},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"487","DOI":"10.1094\/PHYTO-99-5-0487","article-title":"Characterization of sterol demethylation inhibitor-resistant isolates of Fusarium asiaticum and F. graminearum collected from wheat in China","volume":"99","author":"Yin","year":"2009","journal-title":"Phytopathology"},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"839","DOI":"10.3389\/fmicb.2020.00839","article-title":"Population genomic analysis reveals a highly conserved mitochondrial genome in Fusarium asiaticum","volume":"11","author":"Yang","year":"2020","journal-title":"Front Microbiol."},{"key":"ref_9","doi-asserted-by":"crossref","unstructured":"Polak-\u015aliwi\u0144ska, M., and Paszczyk, B. (2021). Trichothecenes in food and feed, relevance to human and animal health and methods of detection: A systematic review. Molecules, 26.","DOI":"10.3390\/molecules26020454"},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"110061","DOI":"10.1016\/j.ijfoodmicro.2022.110061","article-title":"Quantitative PCR assays for the species-specific detection of Fusarium graminearum sensu stricto and Fusarium asiaticum in winter wheat growing regions in China","volume":"387","author":"Wang","year":"2023","journal-title":"Int. J. Food Microbiol."},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"261","DOI":"10.1007\/s13313-017-0487-y","article-title":"Rapid diagnosis of wheat head blight caused by Fusarium asiaticum using a loop-mediated isothermal amplification assay","volume":"46","author":"Xu","year":"2017","journal-title":"Australas Plant Path."},{"key":"ref_12","doi-asserted-by":"crossref","unstructured":"Soroka, M., Wasowicz, B., and Rymaszewska, A. (2021). Loop-mediated isothermal amplification (LAMP): The better sibling of PCR?. Cells, 10.","DOI":"10.3390\/cells10081931"},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"19","DOI":"10.1016\/j.trac.2017.10.015","article-title":"Recombinase polymerase amplification: Basics, applications and recent advances","volume":"98","author":"Lobato","year":"2018","journal-title":"Trends Analyt. Chem."},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"436","DOI":"10.1126\/science.aar6245","article-title":"CRISPR-Cas12a target binding unleashes indiscriminate single-stranded DNase activity","volume":"360","author":"Chen","year":"2018","journal-title":"Science"},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"439","DOI":"10.1126\/science.aaq0179","article-title":"Multiplexed and portable nucleic acid detection platform with Cas13, Cas12a, and Csm6","volume":"360","author":"Gootenberg","year":"2018","journal-title":"Science"},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"20","DOI":"10.1038\/s41421-018-0028-z","article-title":"CRISPR-Cas12a-assisted nucleic acid detection","volume":"4","author":"Li","year":"2018","journal-title":"Cell Discov."},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"1456","DOI":"10.1007\/s11427-021-2028-x","article-title":"A field-deployable method for single and multiplex detection of DNA or RNA from pathogens using Cas12 and Cas13","volume":"65","author":"Li","year":"2022","journal-title":"Sci. China Life Sci."},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"1664","DOI":"10.1094\/PHYTO-03-19-0095-RVW","article-title":"Fusarium head blight in durum wheat: Recent status, breeding directions, and future research prospects","volume":"109","author":"Haile","year":"2019","journal-title":"Phytopathology"},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"eaba5435","DOI":"10.1126\/science.aba5435","article-title":"Horizontal gene transfer of Fhb7 from fungus underlies Fusarium head blight resistance in wheat","volume":"368","author":"Wang","year":"2020","journal-title":"Science"},{"key":"ref_20","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_21","doi-asserted-by":"crossref","unstructured":"Piepenburg, O., Williams, C.H., Stemple, D.L., and Armes, N.A. (2006). DNA Detection Using Recombination Proteins. PLoS Biol., 4.","DOI":"10.1371\/journal.pbio.0040204"},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"e8083","DOI":"10.7717\/peerj.8083","article-title":"A recombinase polymerase amplification-lateral flow dipstick assay for rapid detection of the quarantine citrus pathogen in China, Phytophthora hibernalis","volume":"7","author":"Dai","year":"2019","journal-title":"PeerJ"},{"key":"ref_23","doi-asserted-by":"crossref","unstructured":"Ghosh, D.K., Kokane, S.B., Kokane, A.D., Warghane, A.J., Motghare, M.R., Bhose, S., Sharma, A.K., and Reddy, M.K. (2018). Development of a recombinase polymerase based isothermal amplification combined with lateral flow assay (HLB-RPA-LFA) for rapid detection of \u201cCandidatus Liberibacter asiaticus\u201d. PLoS ONE, 13.","DOI":"10.1371\/journal.pone.0208530"},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"2497","DOI":"10.1021\/acsnano.0c00022","article-title":"Clustered regularly interspaced short palindromic repeats\/Cas9-mediated lateral flow nucleic acid assay","volume":"14","author":"Wang","year":"2020","journal-title":"ACS Nano"},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"421","DOI":"10.1111\/j.1365-2338.2000.tb00922.x","article-title":"On-site detection of plant pathogens using lateral-flow devices","volume":"30","author":"Danks","year":"2000","journal-title":"EPPO Bull."},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"643","DOI":"10.1038\/s41551-021-00760-7","article-title":"CRISPR-based diagnostics","volume":"5","author":"Kaminski","year":"2021","journal-title":"Nat. Biomed. Eng."},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"976510","DOI":"10.3389\/fpls.2022.976510","article-title":"A CRISPR\/Cas12a-based portable platform for rapid detection of Leptosphaeria maculans in Brassica crops","volume":"13","author":"Lei","year":"2022","journal-title":"Front. Plant Sci."},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"2830","DOI":"10.3389\/fmicb.2019.02830","article-title":"Cas12a-based on-site and rapid nucleic acid detection of African Swine Fever","volume":"10","author":"Bai","year":"2019","journal-title":"Front. Microbiol."},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"301","DOI":"10.1016\/j.ijfoodmicro.2010.10.013","article-title":"The CYP51C gene, a reliable marker to resolve interspecific phylogenetic relationships within the Fusarium species complex and a novel target for species-specific PCR","volume":"144","author":"Atkins","year":"2010","journal-title":"Int. J. Food Microbiol."},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"733","DOI":"10.1016\/j.foodcont.2008.09.010","article-title":"Simultaneous detection of Escherichia coli O175:H7, Salmonella spp., and Listeria monocytogenes by multiplex PCR","volume":"20","author":"Germini","year":"2009","journal-title":"Food Control"},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"503","DOI":"10.1080\/00218839.2019.1685148","article-title":"Detection and quantification of Melissococcus plutonius in honey bee workers exposed to European foulbrood in Czechia through conventional PCR, qPCR, and barcode sequencing","volume":"59","author":"Sopko","year":"2020","journal-title":"J. Apicult. Res."},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"183","DOI":"10.1094\/PHYTO-100-2-0183","article-title":"Resistance to Fusarium head blight and seedling blight in wheat is associated with activation of a cytochrome P450 gene","volume":"100","author":"Li","year":"2010","journal-title":"Phytopathology"}],"container-title":["International Journal of Molecular Sciences"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/1422-0067\/24\/18\/14134\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,10]],"date-time":"2025-10-10T20:51:28Z","timestamp":1760129488000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/1422-0067\/24\/18\/14134"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2023,9,15]]},"references-count":32,"journal-issue":{"issue":"18","published-online":{"date-parts":[[2023,9]]}},"alternative-id":["ijms241814134"],"URL":"https:\/\/doi.org\/10.3390\/ijms241814134","relation":{},"ISSN":["1422-0067"],"issn-type":[{"value":"1422-0067","type":"electronic"}],"subject":[],"published":{"date-parts":[[2023,9,15]]}}}