{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,6,16]],"date-time":"2026-06-16T19:57:04Z","timestamp":1781639824462,"version":"3.54.5"},"reference-count":32,"publisher":"Springer Science and Business Media LLC","issue":"1","license":[{"start":{"date-parts":[[2026,3,9]],"date-time":"2026-03-09T00:00:00Z","timestamp":1773014400000},"content-version":"tdm","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0"},{"start":{"date-parts":[[2026,4,15]],"date-time":"2026-04-15T00:00:00Z","timestamp":1776211200000},"content-version":"vor","delay-in-days":37,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0"}],"funder":[{"name":"the University Natural Science Research Key Projects in Anhui Province","award":["2024AH051688, 2024AH051225"],"award-info":[{"award-number":["2024AH051688, 2024AH051225"]}]},{"name":"the University Natural Science Research Key Projects in Anhui Province","award":["2024AH051688, 2024AH051225"],"award-info":[{"award-number":["2024AH051688, 2024AH051225"]}]},{"name":"the University Natural Science Research Key Projects in Anhui Province","award":["2024AH051688, 2024AH051225"],"award-info":[{"award-number":["2024AH051688, 2024AH051225"]}]},{"name":"the University Natural Science Research Key Projects in Anhui Province","award":["2024AH051688, 2024AH051225"],"award-info":[{"award-number":["2024AH051688, 2024AH051225"]}]},{"name":"the University Natural Science Research Key Projects in Anhui Province","award":["2024AH051688, 2024AH051225"],"award-info":[{"award-number":["2024AH051688, 2024AH051225"]}]},{"name":"the University Natural Science Research Key Projects in Anhui Province","award":["2024AH051688, 2024AH051225"],"award-info":[{"award-number":["2024AH051688, 2024AH051225"]}]},{"name":"the University Natural Science Research Key Projects in Anhui Province","award":["2024AH051688, 2024AH051225"],"award-info":[{"award-number":["2024AH051688, 2024AH051225"]}]}],"content-domain":{"domain":["link.springer.com"],"crossmark-restriction":false},"short-container-title":["BMC Microbiol"],"abstract":"<jats:title>Abstract<\/jats:title>\n                  <jats:p>\n                    <jats:italic>Aspergillus fumigatus<\/jats:italic>\n                    (\n                    <jats:italic>AF<\/jats:italic>\n                    ) is the predominant pathogen implicated in invasive aspergillosis (IA) in humans; therefore, prompt and accurate detection is critical for the effective prevention and management of IA. This study developed a rapid detection system targeting the\n                    <jats:italic>AF<\/jats:italic>\n                    -specific\n                    <jats:italic>anxC4<\/jats:italic>\n                    gene by integrating enzymatic recombinase amplification (ERA) with CRISPR\/Cas12a. The reaction proceeds at a stable temperature of 37\u00a0\u00b0C, with amplification and detection systems separately positioned in the tube lid and bottom, respectively, effectively minimizing aerosol contamination typically associated with product transfers. To enhance sensitivity, the One-Pot method was optimized. Consequently, the fluorescence detection limit reached 1\u00a0fg\/\u00b5L, and the sensitivity of the test strip reached 10\u00a0fg\/\u00b5L, with no cross-reactivity observed against other fungi. Detection of\n                    <jats:italic>AF<\/jats:italic>\n                    was completed within 60\u00a0min, and results were visually displayed through fluorescence signals and nucleic acid test strips. Clinical practicality was further evaluated using aspergillosis samples, which demonstrated satisfactory performance. Pure culture results confirmed that out of 62 sputum samples, 32 were positive and 30 negative. Evaluation of 62 clinical samples using the One-Pot ERA-CRISPR\/Cas12a system demonstrated sensitivity and specificity rates of 93.75% and 93.33%, respectively, via fluorescence detection, and 90.63% sensitivity and 96.67% specificity using lateral flow strips.\n                  <\/jats:p>","DOI":"10.1186\/s12866-026-04881-4","type":"journal-article","created":{"date-parts":[[2026,3,9]],"date-time":"2026-03-09T10:05:30Z","timestamp":1773050730000},"update-policy":"https:\/\/doi.org\/10.1007\/springer_crossmark_policy","source":"Crossref","is-referenced-by-count":0,"title":["Development and application of a rapid detection system for Aspergillus fumigatus based on ERA\/CRISPR-Cas12a"],"prefix":"10.1186","volume":"26","author":[{"given":"Qiuyang","family":"Jiang","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Xiaotong","family":"Zeng","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Qi","family":"Zhang","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Fo","family":"Yang","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Tingyao","family":"Lv","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yushuo","family":"Zhang","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jin","family":"Wang","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Feng","family":"Li","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Dayong","family":"Xu","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"297","published-online":{"date-parts":[[2026,3,9]]},"reference":[{"issue":"4","key":"4881_CR1","doi-asserted-by":"publisher","first-page":"57","DOI":"10.3390\/jof3040057","volume":"3","author":"F Bongomin","year":"2017","unstructured":"Bongomin F, Gago S, Oladele RO, Denning DW. Global and multi-national prevalence of fungal diseases estimate precision. J Fungi. 2017;3(4):57.","journal-title":"J Fungi"},{"issue":"7","key":"4881_CR2","doi-asserted-by":"publisher","first-page":"e428","DOI":"10.1016\/S1473-3099(23)00692-8","volume":"24","author":"DW Denning","year":"2024","unstructured":"Denning DW. Global incidence and mortality of severe fungal disease. Lancet Infect Dis. 2024;24(7):e428\u201338.","journal-title":"Lancet Infect Dis"},{"issue":"3","key":"4881_CR3","doi-asserted-by":"publisher","first-page":"312","DOI":"10.1016\/S1473-3099(24)00567-X","volume":"25","author":"A Sengupta","year":"2025","unstructured":"Sengupta A, Ray A, Upadhyay AD, Izumikawa K, Tashiro M, et al. Mortality in chronic pulmonary aspergillosis: a systematic review and individual patient data meta-analysis. Lancet Infect Dis. 2025;25(3):312\u201324.","journal-title":"Lancet Infect Dis"},{"issue":"5","key":"4881_CR4","doi-asserted-by":"publisher","first-page":"2351","DOI":"10.1111\/jam.15092","volume":"131","author":"L Jiang","year":"2021","unstructured":"Jiang L, Gu R, Li X, Mu D. Simple and rapid detection Aspergillus fumigatus by loop-mediated isothermal amplification coupled with lateral flow biosensor assay. J Appl Microbiol. 2021;131(5):2351\u201360.","journal-title":"J Appl Microbiol"},{"issue":"1","key":"4881_CR5","doi-asserted-by":"publisher","first-page":"00140","DOI":"10.1128\/CMR.00140-18","volume":"33","author":"J-P Latg\u00e9","year":"2019","unstructured":"Latg\u00e9 J-P, Chamilos G. Aspergillus fumigatus and Aspergillosis in 2019. Clin Microbiol Rev. 2019;33(1):00140\u201318.","journal-title":"Clin Microbiol Rev"},{"issue":"1","key":"4881_CR6","doi-asserted-by":"publisher","first-page":"105807","DOI":"10.1016\/j.ijantimicag.2019.09.011","volume":"55","author":"A P\u00e9rez-Cantero","year":"2020","unstructured":"P\u00e9rez-Cantero A, L\u00f3pez-Fern\u00e1ndez L, Guarro J, Capilla J. Azole resistance mechanisms in Aspergillus: update and recent advances. Int J Antimicrob Agents. 2020;55(1):105807.","journal-title":"Int J Antimicrob Agents"},{"issue":"5","key":"4881_CR7","doi-asserted-by":"publisher","first-page":"470","DOI":"10.1097\/00043764-200305000-00006","volume":"45","author":"BD Hardin","year":"2003","unstructured":"Hardin BD, Kelman BJ, Saxon A. Adverse human health effects associated with molds in the indoor environment. Int J Occup Environ Med. 2003;45(5):470\u201378.","journal-title":"Int J Occup Environ Med"},{"issue":"3","key":"4881_CR8","doi-asserted-by":"publisher","first-page":"535","DOI":"10.1016\/j.ccm.2017.04.013","volume":"38","author":"P Ramanan","year":"2017","unstructured":"Ramanan P, Wengenack NL, Theel ES. Laboratory diagnostics for fungal infections: a review of current and future diagnostic assays. Clin Chest Med. 2017;38(3):535\u201354.","journal-title":"Clin Chest Med"},{"issue":"4","key":"4881_CR9","doi-asserted-by":"publisher","first-page":"115274","DOI":"10.1016\/j.diagmicrobio.2020.115274","volume":"99","author":"Z Wu","year":"2021","unstructured":"Wu Z, Wang L, Tan L, Wu J, Chen Z, Hu M. Diagnostic value of galactomannan in serum and bronchoalveolar lavage fluid for invasive pulmonary aspergillosis in non-neutropenic patients. Diagn Microbiol Infect Dis. 2021;99(4):115274.","journal-title":"Diagn Microbiol Infect Dis"},{"issue":"12","key":"4881_CR10","doi-asserted-by":"publisher","first-page":"1760","DOI":"10.1128\/CVI.00226-08","volume":"15","author":"S Husain","year":"2008","unstructured":"Husain S, Clancy CJ, Nguyen MH, Swartzentruber S, Leather H, LeMonte AM, et al. Performance characteristics of the platelia Aspergillus enzyme immunoassay for detection of Aspergillus galactomannan antigen in bronchoalveolar lavage fluid. Clin Vaccine Immunol. 2008;15(12):1760\u20133.","journal-title":"Clin Vaccine Immunol"},{"issue":"2","key":"4881_CR11","doi-asserted-by":"publisher","first-page":"286","DOI":"10.1016\/j.jmoldx.2018.10.004","volume":"21","author":"LS Yu","year":"2019","unstructured":"Yu LS, Rodriguez-Manzano J, Malpartida-Cardenas K, Sewell T, Bader O, Armstrong-James D, et al. Rapid and Sensitive Detection of Azole-Resistant Aspergillus fumigatus by Tandem Repeat Loop-Mediated Isothermal Amplification. J Mol Diagn. 2019;21(2):286\u201395.","journal-title":"J Mol Diagn"},{"issue":"3","key":"4881_CR12","doi-asserted-by":"publisher","first-page":"306","DOI":"10.3390\/mi11030306","volume":"11","author":"T Nguyen","year":"2020","unstructured":"Nguyen T, Duong BD, Wolff A. 2019 Novel Coronavirus Disease (COVID-19): paving the road for rapid setection and point-of-care diagnostics. Micromachines (Basel). 2020;11(3):306.","journal-title":"Micromachines (Basel)"},{"key":"4881_CR13","doi-asserted-by":"publisher","first-page":"54","DOI":"10.1016\/j.talanta.2018.08.033","volume":"191","author":"E Shahbazi","year":"2019","unstructured":"Shahbazi E, Mollasalehi H, Minai-Tehrani D. Development and evaluation of an improved quantitative loop-mediated isothermal amplification method for rapid detection of Morganella morganii. Talanta. 2019;191:54\u20138.","journal-title":"Talanta"},{"key":"4881_CR14","doi-asserted-by":"publisher","first-page":"104446","DOI":"10.1016\/j.jcv.2020.104446","volume":"129","author":"Y Kitagawa","year":"2020","unstructured":"Kitagawa Y, Orihara Y, Kawamura R, Imai K, Sakai J, Tarumoto N, et al. Evaluation of rapid diagnosis of novel coronavirus disease (COVID-19) using loop-mediated isothermal amplification. J Clin Virol. 2020;129:104446.","journal-title":"J Clin Virol"},{"issue":"6387","key":"4881_CR15","doi-asserted-by":"publisher","first-page":"436","DOI":"10.1126\/science.aar6245","volume":"360","author":"JS Chen","year":"2018","unstructured":"Chen JS, Ma E, Harrington LB, Da Costa M, Tian X, Palefsky JM, et al. CRISPR-Cas12a target binding unleashes indiscriminate single-stranded DNase activity. Science. 2018;360(6387):436\u20139.","journal-title":"Science"},{"issue":"1","key":"4881_CR16","doi-asserted-by":"publisher","first-page":"20","DOI":"10.1038\/s41421-018-0028-z","volume":"4","author":"SY Li","year":"2018","unstructured":"Li SY, Cheng QX, Wang JM, Li XY, Zhang ZL, Gao S, et al. CRISPR-Cas12a-assisted nucleic acid detection. Cell Discov. 2018;4(1):20.","journal-title":"Cell Discov"},{"key":"4881_CR17","doi-asserted-by":"publisher","first-page":"127608","DOI":"10.1016\/j.foodchem.2020.127608","volume":"334","author":"H Liu","year":"2021","unstructured":"Liu H, Wang J, Zeng H, Liu X, Jiang W, Wang Y, et al. RPA-Cas12a-fs: A frontline nucleic acid rapid detection system for food safety based on CRISPR-Cas12a combined with recombinase polymerase amplification. Food Chem. 2021;334:127608.","journal-title":"Food Chem"},{"issue":"5","key":"4881_CR18","doi-asserted-by":"publisher","first-page":"1427","DOI":"10.1021\/acssensors.0c00320","volume":"5","author":"Y Wang","year":"2020","unstructured":"Wang Y, Ke Y, Liu W, Sun Y, Ding X, et al. A one-pot toolbox based on Cas12a\/crRNA enables rapid foodborne pathogen detection at attomolar level. ACS Sens. 2020;5(5):1427\u201335.","journal-title":"ACS Sens"},{"issue":"11","key":"4881_CR19","doi-asserted-by":"publisher","first-page":"3114","DOI":"10.1021\/acssynbio.0c00365","volume":"9","author":"D Huang","year":"2020","unstructured":"Huang D, Qian J, Shi Z, Zhao J, Fang M, Xu Z, et al. CRISPR-Cas12a-assisted multicolor biosensor for semiquantitative point-of-use testing of the nopaline synthase terminator in genetically modified crops by unaided eyes. ACS Synth Biol. 2020;9(11):3114\u201323.","journal-title":"ACS Synth Biol"},{"issue":"1","key":"4881_CR20","doi-asserted-by":"publisher","first-page":"11","DOI":"10.1007\/s11032-019-1092-2","volume":"40","author":"YM Zhang","year":"2020","unstructured":"Zhang YM, Zhang Y, Xie K. Evaluation of CRISPR\/Cas12a-based DNA detection for fast pathogen diagnosis and GMO test in rice. Mol Breed. 2020;40(1):11.","journal-title":"Mol Breed"},{"issue":"4","key":"4881_CR21","doi-asserted-by":"publisher","first-page":"1587","DOI":"10.1002\/bit.27673","volume":"118","author":"D Huang","year":"2021","unstructured":"Huang D, Shi Z, Qian J, Bi K, Fang M, Xu Z. A CRISPR-Cas12a-derived biosensor enabling portable personal glucose meter readout for quantitative detection of SARS. CoV-2. Biotechnol Bioeng. 2021;118(4):1587\u201396.","journal-title":"Biotechnol Bioeng"},{"issue":"7","key":"4881_CR22","doi-asserted-by":"publisher","first-page":"870","DOI":"10.1038\/s41587-020-0513-4","volume":"38","author":"JP Broughton","year":"2020","unstructured":"Broughton JP, Deng X, Yu G, Fasching CL, Servellita V, Singh J, et al. CRISPR-Cas12-based detection of SARS-CoV-2. Nat Biotechnol. 2020;38(7):870\u20134.","journal-title":"Nat Biotechnol"},{"key":"4881_CR23","doi-asserted-by":"publisher","first-page":"128119","DOI":"10.1016\/j.snb.2020.128119","volume":"316","author":"O Mukama","year":"2020","unstructured":"Mukama O, Yuan T, He Z, Li Z, Zeng L. A high fidelity CRISPR\/Cas12a based lateral flow biosensor for the detection of HPVl6 and HPV18. Sens Actuators B Chem. 2020;316:128119.","journal-title":"Sens Actuators B Chem"},{"key":"4881_CR24","doi-asserted-by":"publisher","first-page":"117458","DOI":"10.1016\/j.bios.2025.117458","volume":"281","author":"J Li","year":"2025","unstructured":"Li J, Huang H, Song Z, Chen S, Xu J, Yang J, et al. Palm-sized CRISPR sensing platform for on-site Mycoplasma pneumoniae detection. Biosens Bioelectron. 2025;281:117458.","journal-title":"Biosens Bioelectron"},{"issue":"1","key":"4881_CR25","doi-asserted-by":"publisher","first-page":"9304","DOI":"10.1038\/s41467-024-53693-5","volume":"15","author":"S Shen","year":"2024","unstructured":"Shen S, Wang W, Ma Y, Wang S, Zhang S, Cai X, et al. Affinity molecular assay for detecting Candida albicans using chitin affinity and RPA-CRISPR\/Cas12a. Nat Commun. 2024;15(1):9304.","journal-title":"Nat Commun"},{"key":"4881_CR26","doi-asserted-by":"publisher","first-page":"198129","DOI":"10.1016\/j.virusres.2020.198129","volume":"288","author":"Z Ali","year":"2020","unstructured":"Ali Z, Aman R, Mahas A, Rao GS, Tehseen M, Marsic T, et al. iSCAN: An RT-LAMP-coupled CRISPR-Cas12 module for rapid, sensitive detection of SARS-CoV-2. Virus Res. 2020;288:198129.","journal-title":"Virus Res"},{"issue":"12","key":"4881_CR27","doi-asserted-by":"publisher","first-page":"8561","DOI":"10.1021\/acs.analchem.0c01459","volume":"92","author":"K Yin","year":"2020","unstructured":"Yin K, Ding X, Li Z, Zhao H, Cooper K, Liu C. Dynamic Aqueous Multiphase Reaction System for One-Pot CRISPR-Cas12a-Based Ultrasensitive and Quantitative Molecular Diagnosis. Anal Chem. 2020;92(12):8561\u20138.","journal-title":"Anal Chem"},{"issue":"1\u20133","key":"4881_CR28","doi-asserted-by":"publisher","first-page":"79","DOI":"10.1016\/S0014-5793(04)00186-3","volume":"562","author":"V Khalaj","year":"2004","unstructured":"Khalaj V, Smith L, Brookman J, Tuckwell D. Identification of a novel class of annexin genes. FEBS Lett. 2004;562(1\u20133):79\u201386.","journal-title":"FEBS Lett"},{"issue":"23","key":"4881_CR29","doi-asserted-by":"publisher","first-page":"8277","DOI":"10.1021\/acs.analchem.2c00616","volume":"94","author":"M Lin","year":"2022","unstructured":"Lin M, Yue H, Tian T, Xiong E, Zhu D, Jiang Y, et al. Glycerol Additive Boosts 100-fold Sensitivity Enhancement for One-Pot RPA-CRISPR\/Cas12a Assay. Anal Chem. 2022;94(23):8277\u201384.","journal-title":"Anal Chem"},{"issue":"3","key":"4881_CR30","doi-asserted-by":"publisher","first-page":"1381","DOI":"10.1021\/acs.jafc.2c06549","volume":"71","author":"JJ Hu","year":"2023","unstructured":"Hu JJ, Liu D, Cai MZ, Zhou Y, Yin WX, Luo CX. One-Pot Assay for Rapid Detection of Benzimidazole Resistance in Venturia carpophila by Combining RPA and CRISPR\/Cas12a. J Agric Food Chem. 2023;71(3):1381\u201390.","journal-title":"J Agric Food Chem"},{"issue":"4","key":"4881_CR31","doi-asserted-by":"publisher","first-page":"749","DOI":"10.1016\/j.jmii.2021.11.008","volume":"55","author":"Z Li","year":"2022","unstructured":"Li Z, Wang M, Xu T, et al. Development and clinical implications of a novel CRISPR-based diagnostic test for pulmonary Aspergillus fumigatus infection. J Microbiol Immunol Infect. 2022;55(4):749\u201356.","journal-title":"J Microbiol Immunol Infect"},{"issue":"4","key":"4881_CR32","doi-asserted-by":"publisher","first-page":"116","DOI":"10.1007\/s11274-024-03940-0","volume":"40","author":"C Lin","year":"2024","unstructured":"Lin C, Zhou J, Gao N, et al. Establishing a pulmonary Aspergillus fumigatus infection diagnostic platform based on RPA-CRISPR-Cas12a. World J Microbiol Biotechnol. 2024;40(4):116.","journal-title":"World J Microbiol Biotechnol"}],"container-title":["BMC Microbiology"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/link.springer.com\/article\/10.1186\/s12866-026-04881-4","content-type":"text\/html","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/link.springer.com\/content\/pdf\/10.1186\/s12866-026-04881-4.pdf","content-type":"application\/pdf","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/link.springer.com\/content\/pdf\/10.1186\/s12866-026-04881-4.pdf","content-type":"application\/pdf","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2026,6,16]],"date-time":"2026-06-16T19:37:38Z","timestamp":1781638658000},"score":1,"resource":{"primary":{"URL":"https:\/\/link.springer.com\/10.1186\/s12866-026-04881-4"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2026,3,9]]},"references-count":32,"journal-issue":{"issue":"1","published-online":{"date-parts":[[2026,12]]}},"alternative-id":["4881"],"URL":"https:\/\/doi.org\/10.1186\/s12866-026-04881-4","relation":{"has-preprint":[{"id-type":"doi","id":"10.21203\/rs.3.rs-7538348\/v1","asserted-by":"object"}]},"ISSN":["1471-2180"],"issn-type":[{"value":"1471-2180","type":"electronic"}],"subject":[],"published":{"date-parts":[[2026,3,9]]},"assertion":[{"value":"4 September 2025","order":1,"name":"received","label":"Received","group":{"name":"ArticleHistory","label":"Article History"}},{"value":"19 February 2026","order":2,"name":"accepted","label":"Accepted","group":{"name":"ArticleHistory","label":"Article History"}},{"value":"9 March 2026","order":3,"name":"first_online","label":"First Online","group":{"name":"ArticleHistory","label":"Article History"}},{"order":1,"name":"Ethics","group":{"name":"EthicsHeading","label":"Declarations"}},{"value":"The studies involving clinical samples were reviewed and approved by the Huaibei. People\u2019s Hospital Ethics Committee (no. 2024\u2013052). Informed consent was obtained from all participants involved in the study. The study was in compliance with the Declaration of Helsinki.","order":2,"name":"Ethics","group":{"name":"EthicsHeading","label":"Ethics approval and consent to participate"}},{"value":"Not applicable.","order":3,"name":"Ethics","group":{"name":"EthicsHeading","label":"Consent for publication"}},{"value":"The authors declare no competing interests.","order":4,"name":"Ethics","group":{"name":"EthicsHeading","label":"Competing interests"}}],"article-number":"359"}}