{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,8,20]],"date-time":"2026-08-20T05:24:00Z","timestamp":1787203440817,"version":"build-2736575974"},"reference-count":57,"publisher":"MDPI AG","issue":"8","license":[{"start":{"date-parts":[[2026,8,18]],"date-time":"2026-08-18T00:00:00Z","timestamp":1787011200000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"Xinjiang Uygur Autonomous Region key research and development task special","award":["2024B02014-2"],"award-info":[{"award-number":["2024B02014-2"]}]},{"name":"National Key Research and Development Program of China","award":["2023YFD2400704"],"award-info":[{"award-number":["2023YFD2400704"]}]},{"name":"Central Public-interest Scientific Institution Basal Research Fund","award":["2023TD46"],"award-info":[{"award-number":["2023TD46"]}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Vet. Sci.","Veterinary Sciences"],"abstract":"<jats:p>The high mortality associated with largemouth bass ranavirus (LMBRaV) disease requires rapid and sensitive diagnosis methods to prevent virus spread and subsequent outbreak. The recombinase polymerase amplification combined with the CRISPR\/Cas12a system (RPA-CRISPR\/Cas12a) assay and RPA-CRISPR\/Cas12a, coupled with lateral flow dipstick (RPA-CRISPR\/Cas12a-LFD) assay for LMBRaV, were established in this study for low viral load surveillance. The major capsid protein (mcp) gene is highly conserved and is widely used as the target sequence in LMBRaV detection. First, based on the mcp gene sequence of LMBRaV, three candidate crRNAs were designed. Among them, the crRNA-2, which exhibited the highest cleavage activity, was selected through dual evaluation of fluorescence signal intensity and LFD color development. Then, the key reaction conditions for both assays were optimized as follows: LbCas12a protein concentration of 150 nM, crRNA-2 concentration of 200 nM, and ssDNA reporters\u2019 concentration of 200 nM. Moreover, specificity evaluation showed that both combined assays specifically recognized LMBRaV, with no cross-reactivity detected against other aquatic pathogens, such as CyHV-2, GSIV, ISKNV, WSSV, GCRV II or CrERV. The detection limit was 1 copy\/\u03bcL of DNA sample for both RPA-CRISPR\/Cas12a and RPA-CRISPR\/Cas12a-LFD assays. Finally, validation using 24 clinical samples (16 positive, eight negative) showed that both LMBRaV RPA-CRISPR\/Cas12a assays achieved 100% detection rate, whereas conventional PCR detected 13 positive samples (detection rate 81.26%). ddPCR served as the reference method for clinical validation. Therefore, the LMBRaV RPA-CRISPR\/Cas12a assay and the RPA-CRISPR\/Cas12a-LFD assay provide sensitive, specific, and easy-to-operate methods for the rapid detection of LMBRaV.<\/jats:p>","DOI":"10.3390\/vetsci13080825","type":"journal-article","created":{"date-parts":[[2026,8,18]],"date-time":"2026-08-18T12:09:32Z","timestamp":1787054972000},"page":"825","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":0,"title":["Establishment of RPA-CRISPR\/Cas12a Detection Methods for Rapid Largemouth Bass Ranavirus Surveillance"],"prefix":"10.3390","volume":"13","author":[{"given":"Haoyu","family":"Wang","sequence":"first","affiliation":[{"name":"Division of Fish Disease, Yangtze River Fisheries Research Institute, Chinese Academy of Fishery Sciences, Wuhan 430223, China"},{"name":"Engineering Lab of Henan Province for Aquatic Animal Disease Control, College of Fisheries, Henan Normal University, Xinxiang 453007, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-0373-5734","authenticated-orcid":false,"given":"Yong","family":"Zhou","sequence":"additional","affiliation":[{"name":"Division of Fish Disease, Yangtze River Fisheries Research Institute, Chinese Academy of Fishery Sciences, Wuhan 430223, China"},{"name":"Engineering Lab of Henan Province for Aquatic Animal Disease Control, College of Fisheries, Henan Normal University, Xinxiang 453007, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Peng","family":"Chen","sequence":"additional","affiliation":[{"name":"Xinjiang Uygur Autonomous Region Fisheries Development Center (Xinjiang Uygur Autonomous Region Fisheries Research Institute), Urumqi 830000, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Liping","family":"Zhang","sequence":"additional","affiliation":[{"name":"Chongqing Fisheries Technical Extension Center, Chongqing 401120, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Wen","family":"Zhu","sequence":"additional","affiliation":[{"name":"Division of Fish Disease, Yangtze River Fisheries Research Institute, Chinese Academy of Fishery Sciences, Wuhan 430223, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-3370-5712","authenticated-orcid":false,"given":"Mingyang","family":"Xue","sequence":"additional","affiliation":[{"name":"Division of Fish Disease, Yangtze River Fisheries Research Institute, Chinese Academy of Fishery Sciences, Wuhan 430223, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-8846-2191","authenticated-orcid":false,"given":"Yan","family":"Meng","sequence":"additional","affiliation":[{"name":"Division of Fish Disease, Yangtze River Fisheries Research Institute, Chinese Academy of Fishery Sciences, Wuhan 430223, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Zhenyu","family":"Huang","sequence":"additional","affiliation":[{"name":"Division of Fish Disease, Yangtze River Fisheries Research Institute, Chinese Academy of Fishery Sciences, Wuhan 430223, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-5075-7154","authenticated-orcid":false,"given":"Chen","family":"Xu","sequence":"additional","affiliation":[{"name":"Division of Fish Disease, Yangtze River Fisheries Research Institute, Chinese Academy of Fishery Sciences, Wuhan 430223, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-7291-2493","authenticated-orcid":false,"given":"Yuding","family":"Fan","sequence":"additional","affiliation":[{"name":"Division of Fish Disease, Yangtze River Fisheries Research Institute, Chinese Academy of Fishery Sciences, Wuhan 430223, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Chao","family":"Pei","sequence":"additional","affiliation":[{"name":"Engineering Lab of Henan Province for Aquatic Animal Disease Control, College of Fisheries, Henan Normal University, Xinxiang 453007, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-8460-3949","authenticated-orcid":false,"given":"Nan","family":"Jiang","sequence":"additional","affiliation":[{"name":"Division of Fish Disease, Yangtze River Fisheries Research Institute, Chinese Academy of Fishery Sciences, Wuhan 430223, China"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2026,8,18]]},"reference":[{"key":"ref_1","first-page":"73","article-title":"Progress on viral disease caused by largemouth bass ranavirus","volume":"32","author":"Wang","year":"2011","journal-title":"Prog. Vet. Med."},{"key":"ref_2","unstructured":"Agriculture and Rural Affairs Ministry Fisheries Administration Bureau (2025). China Fishery Statistical Yearbook 2025, Agricultural Press of China."},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"e70016","DOI":"10.1111\/jfd.70016","article-title":"Establishment of basic-RPA and RPA-LFD rapid detection methods for ranavirus in largemouth bass (Micropterus salmoides)","volume":"49","author":"Wang","year":"2026","journal-title":"J. Fish. Dis."},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"198","DOI":"10.1016\/j.aquaculture.2010.12.032","article-title":"Characterization of a ranavirus isolated from cultured largemouth bass (Micropterus salmoides) in China","volume":"312","author":"Deng","year":"2011","journal-title":"Aquaculture"},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"265","DOI":"10.1577\/1548-8667(1996)008<0265:AIIFWL>2.3.CO;2","article-title":"An iridovirus isolated from wild largemouth bass","volume":"8","author":"Plumb","year":"1996","journal-title":"J. Aquat. Anim. Health"},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"233","DOI":"10.3354\/dao050233","article-title":"First isolation of largemouth bass virus","volume":"50","author":"Grizzle","year":"2002","journal-title":"Dis. Aquat. Org."},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"10","DOI":"10.1577\/1548-8446(2003)28[10:ROLBV]2.0.CO;2","article-title":"Review of largemouth bass virus","volume":"28","author":"Grizzle","year":"2003","journal-title":"Fisheries"},{"key":"ref_8","first-page":"871","article-title":"Isolation and preliminary identification of the pathogen from largemouth bass ulcerative syndrome","volume":"33","author":"Deng","year":"2009","journal-title":"J. Fish. China"},{"key":"ref_9","first-page":"156","article-title":"Characterization of an iridovirus isolate from largemouth bass Micropterus salmoides","volume":"51","author":"Xu","year":"2020","journal-title":"Oceanol. Et Limnol. Sin."},{"key":"ref_10","first-page":"494","article-title":"Isolation and identification of largemouth bass ranavirus strain LMBRaV-HB001 isolated from diseased Micropterus salmoides in Hubei Province, China","volume":"29","author":"Luo","year":"2022","journal-title":"J. Fish. Sci. China"},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"95","DOI":"10.1016\/j.exppara.2016.03.020","article-title":"Antiparasitic effects of oxymatrine and matrine against Toxoplasma gondii in vitro and in vivo","volume":"165","author":"Zhang","year":"2016","journal-title":"Exp. Parasitol."},{"key":"ref_12","first-page":"508","article-title":"Establishment and application of TaqMan qPCR method for largemouth bass ranavirus","volume":"44","author":"Gong","year":"2022","journal-title":"Chin. J. Prev. Vet. Med."},{"key":"ref_13","first-page":"146","article-title":"Dynamic pathological lesions and tissue distribution of Chinese giant salamanders infected with CGSRV","volume":"24","author":"Liu","year":"2017","journal-title":"J. Fish. China"},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"143","DOI":"10.3354\/dao039143","article-title":"Preliminary description of lesions in juvenile largemouth bass injected with largemouth bass virus","volume":"39","author":"Zilberg","year":"2000","journal-title":"Dis. Aquat. Org."},{"key":"ref_15","first-page":"623","article-title":"Effect of in vitro passage on virulence of largemouth bass (Micropterus salmoides) ranavirus","volume":"38","author":"Wang","year":"2023","journal-title":"J. Dalian Ocean Univ."},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"108641","DOI":"10.1016\/j.fsi.2023.108641","article-title":"Isolation, identification and the pathogenicity characterization of a santee-cooper ranavirus and its activation on immune responses in juvenile largemouth bass (Micropterus salmoides)","volume":"135","author":"Liu","year":"2023","journal-title":"Fish. Shellfish Immunol."},{"key":"ref_17","first-page":"062503","article-title":"Evaluation of immune effect on attenuated strain of largemouth bass ranavirus","volume":"49","author":"Gao","year":"2025","journal-title":"Acta Hydrobiol. Sin."},{"key":"ref_18","first-page":"184","article-title":"Isolation, identification and histopathology of largemouth bass ranavirus (Iridoviridae) from Micropterus salmoides","volume":"55","author":"Li","year":"2025","journal-title":"Period. Ocean Univ. China"},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"389","DOI":"10.1111\/jfd.12246","article-title":"Isolation and characterization of a ranavirus from koi, Cyprinus carpio L., experiencing mass mortalities in India","volume":"38","author":"George","year":"2015","journal-title":"J. Fish. Dis."},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"29","DOI":"10.3354\/dao054029","article-title":"PCR method for detection of largemouth bass virus","volume":"54","author":"Grizzle","year":"2003","journal-title":"Dis. Aquat. Org."},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"101569","DOI":"10.1016\/j.mcp.2020.101569","article-title":"Rapid diagnosis of largemouth bass ranavirus in fish samples using the loop-mediated isothermal amplification method","volume":"52","author":"Zhu","year":"2020","journal-title":"Mol. Cell. Probes"},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"226","DOI":"10.1577\/H07-009.1","article-title":"Quantitative polymerase chain reaction assay for largemouth bass virus","volume":"19","author":"Getchell","year":"2007","journal-title":"J. Aquat. Anim. Health"},{"key":"ref_23","doi-asserted-by":"crossref","unstructured":"Lennemann, N.J., and Coyne, C.B. (2015). Catch me if you can: The link between autophagy and viruses. PLoS Pathog., 11.","DOI":"10.1371\/journal.ppat.1004685"},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"91","DOI":"10.1111\/jfd.13721","article-title":"Development of a droplet digital PCR method for the sensitive detection and quantification of largemouth bass ranavirus","volume":"46","author":"Jiang","year":"2023","journal-title":"J. Fish. Dis."},{"key":"ref_25","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_26","doi-asserted-by":"crossref","first-page":"737907","DOI":"10.1016\/j.aquaculture.2022.737907","article-title":"Onsite and visual detection of ranavirus in largemouth bass (Micropterus salmoides) by an isothermal recombinase polymerase amplification method","volume":"551","author":"Zhu","year":"2022","journal-title":"Aquaculture"},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"759","DOI":"10.1016\/j.cell.2015.09.038","article-title":"Cpf1 is a single RNA-guided endonuclease of a class 2 CRISPR-cas system","volume":"163","author":"Zetsche","year":"2015","journal-title":"Cell"},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"491","DOI":"10.1038\/s41422-018-0022-x","article-title":"CRISPR-Cas12a has both cis- and trans-cleavage activities on single-stranded DNA","volume":"28","author":"Li","year":"2018","journal-title":"Cell Res."},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"38","DOI":"10.1016\/j.jviromet.2017.12.008","article-title":"Development of cross-priming amplification coupled with vertical flow visualization for rapid detection of infectious spleen and kidney necrosis virus (ISKNV) in mandarin fish, Siniperca chuatsi","volume":"253","author":"Liu","year":"2018","journal-title":"J. Virol. Methods"},{"key":"ref_30","doi-asserted-by":"crossref","unstructured":"Lin, Z., Lu, J., Wu, S., Lin, X., Zheng, L., Lou, Y., and Xiao, X. (2022). A novel detection method for the pathogenic Aeromonas hydrophila expressing aerA gene and\/or hlyA gene based on dualplex RAA and CRISPR\/Cas12a. Front. Microbiol., 13.","DOI":"10.3389\/fmicb.2022.973996"},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"125967","DOI":"10.1016\/j.ijbiomac.2023.125967","article-title":"Rapid, sensitive, and visual detection of Clonorchis sinensis with an RPA-CRISPR\/Cas12a-based dual readout portable platform","volume":"249","author":"Huang","year":"2023","journal-title":"Int. J. Biol. Macromole."},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"343469","DOI":"10.1016\/j.aca.2024.343469","article-title":"Dual-CRISPR\/Cas12a-assisted RT-RAA visualization system for rapid on-site detection of nervous necrosis virus (NNV)","volume":"1335","author":"Gao","year":"2025","journal-title":"Anal. Chim. Acta"},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"e14059","DOI":"10.1111\/jfd.14059","article-title":"Multiplex CRISPR-cas assay for rapid, isothermal and visual detection of white spot syndrome virus (WSSV) and enterocytozoon hepatopenaei (EHP) in penaeid shrimp","volume":"48","author":"Kanitchinda","year":"2025","journal-title":"J. Fish. Dis."},{"key":"ref_34","doi-asserted-by":"crossref","unstructured":"Lu, Z., Liang, M., Li, C., Xu, Y., Weng, S., He, J., and Guo, C. (2024). Rapid, sensitive, and visual detection of mandarin fish ranavirus and infectious spleen and kidney necrosis virus using an RPA-CRISPR\/Cas12a system. Front. Microbiol., 15.","DOI":"10.3389\/fmicb.2024.1495777"},{"key":"ref_35","doi-asserted-by":"crossref","first-page":"740658","DOI":"10.1016\/j.aquaculture.2024.740658","article-title":"Development and validation of a CRISPR\/Cas12a-based platform for rapid and sensitive Detection of the large yellow croaker iridovirus","volume":"584","author":"Zhang","year":"2024","journal-title":"Aquaculture"},{"key":"ref_36","doi-asserted-by":"crossref","first-page":"742704","DOI":"10.1016\/j.aquaculture.2025.742704","article-title":"Development of a rapid and visual detection platform for Pseudomonas plecoglossicida in large yellow croaker (Larimichthys crocea) based on the RPA-CRISPR\/Cas12a system","volume":"608","author":"Jiang","year":"2025","journal-title":"Aquaculture"},{"key":"ref_37","doi-asserted-by":"crossref","first-page":"e70095","DOI":"10.1111\/jfd.70095","article-title":"Development of an RPA-CRISPR-Cas12a fluorescence assay for rapid and sensitive detection of Tilapia parvovirus (TiPV)","volume":"49","author":"Ma","year":"2026","journal-title":"J. Fish. Dis."},{"key":"ref_38","doi-asserted-by":"crossref","first-page":"170","DOI":"10.1186\/s12985-026-03185-3","article-title":"Establishment and application of a detection method for Chinese rice-field eels rhabdovirus (CrERV) using the RPA- CRISPR\/Cas12a system","volume":"23","author":"Su","year":"2026","journal-title":"Virol. J."},{"key":"ref_39","doi-asserted-by":"crossref","unstructured":"Li, N., Dai, C., Mao, Y., Zhang, Y., Yan, H., Wang, H., Qin, Y., Huang, B., Wang, X., and Yao, L. (2025). Development of a rapid on-site detection method for largemouth bass virus based on RPA-CRISPR\/Cas12a system. Front. Microbiol., 16.","DOI":"10.3389\/fmicb.2025.1599006"},{"key":"ref_40","doi-asserted-by":"crossref","first-page":"211","DOI":"10.1016\/j.jviromet.2013.08.024","article-title":"Development of a loop-mediated isothermal amplification assay for rapid detection of iridovirus in the chinese giant salamander","volume":"194","author":"Meng","year":"2013","journal-title":"J. Virol. Methods"},{"key":"ref_41","doi-asserted-by":"crossref","first-page":"138","DOI":"10.1016\/j.vetmic.2013.05.025","article-title":"Cyprinid herpesvirus 2 infection emerged in cultured gibel carp, Carassius auratus gibelio in China","volume":"166","author":"Xu","year":"2013","journal-title":"Vet. Microbiol."},{"key":"ref_42","first-page":"1149","article-title":"Sequence analysis of MCP gene from largemouth bass ulcerative syndrome virus and rapid detection by PCR assay","volume":"17","author":"Ma","year":"2010","journal-title":"J. Fish. China"},{"key":"ref_43","doi-asserted-by":"crossref","first-page":"148595","DOI":"10.1016\/j.foodchem.2026.148595","article-title":"crRNA-engineered CRISPR\/Cas12a system coupled with RPA for ultrasensitive detection of Lactiplantibacillus plantarum","volume":"509","author":"Yu","year":"2026","journal-title":"Food Chem."},{"key":"ref_44","doi-asserted-by":"crossref","first-page":"e04233-22","DOI":"10.1128\/spectrum.04233-22","article-title":"CRISPR\/CAS12A technology combined with recombinase polymerase amplification for rapid and portable monkeypox virus detection","volume":"11","author":"Li","year":"2023","journal-title":"Microbiol. Spectr."},{"key":"ref_45","doi-asserted-by":"crossref","first-page":"6490","DOI":"10.1021\/acs.jafc.2c08689","article-title":"One-pot molecular diagnosis of acute hepatopancreatic necrosis disease by recombinase polymerase amplification and CRISPR\/Cas12a with specially designed crRNA","volume":"71","author":"Wang","year":"2023","journal-title":"J. Agric. Food Chem."},{"key":"ref_46","doi-asserted-by":"crossref","unstructured":"Muriuki, R., Ndichu, M., Githigia, S., and Svitek, N. (2024). CRISPR-cas based Pen-Side diagnostic tests for Anaplasma marginale and Babesia bigemina. Microorganisms, 12.","DOI":"10.3390\/microorganisms12122595"},{"key":"ref_47","doi-asserted-by":"crossref","unstructured":"Tan, M., Liao, C., Liang, L., Yi, X., Zhou, Z., and Wei, G. (2022). Recent advances in recombinase polymerase amplification: Principle, advantages, disadvantages and applications. Front. Cell. Infect. Microbiol., 12.","DOI":"10.3389\/fcimb.2022.1019071"},{"key":"ref_48","doi-asserted-by":"crossref","first-page":"9724","DOI":"10.1021\/acs.analchem.2c01193","article-title":"Photoactivatable CRISPR\/Cas12a strategy for one-pot DETECTR molecular diagnosis","volume":"94","author":"Chen","year":"2022","journal-title":"Anal. Chem."},{"key":"ref_49","doi-asserted-by":"crossref","first-page":"10443","DOI":"10.1021\/acs.analchem.4c01777","article-title":"In situ complexation of sgRNA and Cas12a improves the performance of a one-pot RPA\u2013CRISPR-Cas12 assay","volume":"96","author":"Lesinski","year":"2024","journal-title":"Anal. Chem."},{"key":"ref_50","doi-asserted-by":"crossref","first-page":"148872","DOI":"10.1016\/j.cej.2024.148872","article-title":"Spatiotemporal dual-dimensional separation enables ultra-simple one-pot CRISPR-based molecular diagnostics","volume":"482","author":"Li","year":"2024","journal-title":"Chem. Eng. J."},{"key":"ref_51","doi-asserted-by":"crossref","first-page":"168126","DOI":"10.1016\/j.cej.2025.168126","article-title":"Highly responsive push-orchestrated-pull switching-enabled RNA-regulated Cas12a cis-cleavage: A universal one-pot multiplex-targeted detection platform with application to Mycobacterium tuberculosis complex","volume":"523","author":"Yang","year":"2025","journal-title":"Chem. Eng. J."},{"key":"ref_52","doi-asserted-by":"crossref","unstructured":"Fu, J., Zhang, Y., Cai, G., Meng, G., and Shi, S. (2021). Rapid and sensitive RPA-Cas12a-fluorescence assay for point-of-care detection of African swine fever virus. PLoS ONE, 16.","DOI":"10.1371\/journal.pone.0254815"},{"key":"ref_53","doi-asserted-by":"crossref","unstructured":"Wen, S.B., She, L.M.G., Dang, S., Liao, A., Li, X., Zhang, S., Song, Y., Li, X., and Zhai, J. (2024). Development of a RPACRISPR\/Cas12a based rapid visual detection assay for Porcine Parvovirus 7. Front. Vet. Sci., 11.","DOI":"10.3389\/fvets.2024.1440769"},{"key":"ref_54","doi-asserted-by":"crossref","first-page":"e202300663","DOI":"10.1002\/anie.202300663","article-title":"Light-start CRISPR-Cas12a reaction with caged crRNA enables rapid and sensitive nucleic acid detection","volume":"62","author":"Hu","year":"2023","journal-title":"Angew. Chem."},{"key":"ref_55","doi-asserted-by":"crossref","first-page":"1922","DOI":"10.1016\/j.scib.2024.12.032","article-title":"Heat-triggered CRISPR\/Cas12a microspheres for one-pot assays","volume":"70","author":"Xie","year":"2025","journal-title":"Sci. Bull."},{"key":"ref_56","doi-asserted-by":"crossref","first-page":"169704","DOI":"10.1016\/j.cej.2025.169704","article-title":"Breaking the PAM barrier: CRISPR-Cas12a-based ultrasensitive detection of single-nucleotide variations","volume":"525","author":"Ai","year":"2025","journal-title":"Chem. Eng. J."},{"key":"ref_57","doi-asserted-by":"crossref","first-page":"159693","DOI":"10.1016\/j.cej.2025.159693","article-title":"High performance piperidinium based anion exchange membranes with different twisted backbone structures for fuel cells","volume":"505","author":"Liu","year":"2025","journal-title":"Chem. Eng. J."}],"container-title":["Veterinary Sciences"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/2306-7381\/13\/8\/825\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2026,8,20]],"date-time":"2026-08-20T04:33:26Z","timestamp":1787200406000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/2306-7381\/13\/8\/825"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2026,8,18]]},"references-count":57,"journal-issue":{"issue":"8","published-online":{"date-parts":[[2026,8]]}},"alternative-id":["vetsci13080825"],"URL":"https:\/\/doi.org\/10.3390\/vetsci13080825","relation":{},"ISSN":["2306-7381"],"issn-type":[{"value":"2306-7381","type":"electronic"}],"subject":[],"published":{"date-parts":[[2026,8,18]]}}}