{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,4,19]],"date-time":"2026-04-19T05:30:25Z","timestamp":1776576625963,"version":"3.51.2"},"reference-count":42,"publisher":"American Society for Microbiology","issue":"1","license":[{"start":{"date-parts":[[2024,1,11]],"date-time":"2024-01-11T00:00:00Z","timestamp":1704931200000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"},{"start":{"date-parts":[[2024,1,11]],"date-time":"2024-01-11T00:00:00Z","timestamp":1704931200000},"content-version":"tdm","delay-in-days":0,"URL":"https:\/\/journals.asm.org\/non-commercial-tdm-license"}],"funder":[{"name":"MOST | National Key Research and Development Program of China","award":["2022YFD1800100, 2019YFE0103900"],"award-info":[{"award-number":["2022YFD1800100, 2019YFE0103900"]}]},{"DOI":"10.13039\/501100001809","name":"MOST | National Natural Science Foundation of China","doi-asserted-by":"crossref","award":["32102620"],"award-info":[{"award-number":["32102620"]}],"id":[{"id":"10.13039\/501100001809","id-type":"DOI","asserted-by":"crossref"}]},{"DOI":"10.13039\/501100001809","name":"MOST | National Natural Science Foundation of China","doi-asserted-by":"crossref","award":["31872837"],"award-info":[{"award-number":["31872837"]}],"id":[{"id":"10.13039\/501100001809","id-type":"DOI","asserted-by":"crossref"}]},{"name":"MOST | NSFC | NSFC-Zhejiang Joint Fund | \u6d59\u6c5f\u7701\u79d1\u5b66\u6280\u672f\u5385 | Natural Science Foundation of Zhejiang Province","award":["LZ22C180004"],"award-info":[{"award-number":["LZ22C180004"]}]},{"name":"Joint Project of Hainan Provincial Science and Technology Program and Sanya Yazhou Bay Science and Technology","award":["2021JJLH0083"],"award-info":[{"award-number":["2021JJLH0083"]}]}],"content-domain":{"domain":["journals.asm.org"],"crossmark-restriction":true},"short-container-title":["Microbiol Spectr"],"published-print":{"date-parts":[[2024,1,11]]},"abstract":"<jats:title>ABSTRACT<\/jats:title>\n          <jats:p>Pseudorabies virus (PRV) is the causative pathogen of Aujeszky\u2019s disease. It causes high mortality and miscarriage rates in the infected swine, leading to tremendous economic losses in the global swine industry. PRV has also been demonstrated to trigger viral encephalitis in humans. The eradication policy and large-scale vaccination have been adopted globally as the most effective strategy against PRV. A simple, fast, and sensitive diagnostic method is highly demanded to differentiate between vaccinated and infected swine. Herein, we designed a detection system combining multienzyme isothermal rapid amplification (MIRA) and CRISPR-Cas12a (termed MIRA-Cas12a), characterized by high sensitivity and specificity, low cost, less equipment, and convenient visualization. By targeting the gB, gE, and TK genes of PRV, the MIRA-Cas12a assay is able to distinguish the infected, uninfected, and vaccinated swine by the naked eyes in 40 min (from DNA extraction to result readout) and with comparable sensitivity to conventional quantitative PCR. A 37\u00b0C heater and a source of blue light are all the equipment required to detect PRV. Thus, the MIRA-Cas12a detection will facilitate PRV surveillance and minimize the financial losses to the swine industry.<\/jats:p>\n          <jats:sec>\n            <jats:title>IMPORTANCE<\/jats:title>\n            <jats:p>Pseudorabies virus (PRV) causes high mortality and miscarriage rates in the infected swine, and the eradication policy coupled with large-scale vaccination of live attenuated vaccines has been adopted globally against PRV. Differential diagnosis of the vaccinated and infected swine is highly demanded. Our multienzyme isothermal rapid amplification (MIRA)-Cas12a detection method described in this study can diagnose PRV with a superior sensitivity comparable to the quantitative PCR (qPCR) and a competitive detection speed (only half the time as qPCR needs). The portable feature and the simple procedure of MIRA-Cas12a make it easier to deploy for clinical diagnosis, even in resource-limited settings. The MIRA-Cas12a method would provide immediate and accurate diagnostic information for policymakers to respond promptly.<\/jats:p>\n          <\/jats:sec>","DOI":"10.1128\/spectrum.02617-23","type":"journal-article","created":{"date-parts":[[2023,12,11]],"date-time":"2023-12-11T14:00:34Z","timestamp":1702303234000},"update-policy":"https:\/\/doi.org\/10.1128\/asmj-crossmark-policy-page","source":"Crossref","is-referenced-by-count":14,"title":["Fast and sensitive differential diagnosis of pseudorabies virus-infected versus pseudorabies virus-vaccinated swine using CRISPR-Cas12a"],"prefix":"10.1128","volume":"12","author":[{"ORCID":"https:\/\/orcid.org\/0000-0001-6985-4643","authenticated-orcid":false,"given":"Hao","family":"Wang","sequence":"first","affiliation":[{"name":"Department of Veterinary Medicine, Institute of Preventive Veterinary Sciences, Zhejiang University College of Animal Sciences , Hangzhou, Zhejiang, China"},{"name":"Hainan Institute of Zhejiang University , Sanya, Hainan, China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Hongzhao","family":"Li","sequence":"additional","affiliation":[{"name":"Department of Veterinary Medicine, Institute of Preventive Veterinary Sciences, Zhejiang University College of Animal Sciences , Hangzhou, Zhejiang, China"},{"name":"Hainan Institute of Zhejiang University , Sanya, Hainan, China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Bo","family":"Tang","sequence":"additional","affiliation":[{"name":"Institute of Veterinary Immunology and Engineering, Jiangsu Academy of Agricultural Sciences , Nanjing, Jiangsu, China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Chen","family":"Ye","sequence":"additional","affiliation":[{"name":"Department of Veterinary Medicine, Institute of Preventive Veterinary Sciences, Zhejiang University College of Animal Sciences , Hangzhou, Zhejiang, China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Meiqing","family":"Han","sequence":"additional","affiliation":[{"name":"Department of Veterinary Medicine, Institute of Preventive Veterinary Sciences, Zhejiang University College of Animal Sciences , Hangzhou, Zhejiang, China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Lin","family":"Teng","sequence":"additional","affiliation":[{"name":"Department of Veterinary Medicine, Institute of Preventive Veterinary Sciences, Zhejiang University College of Animal Sciences , Hangzhou, Zhejiang, China"},{"name":"Zhejiang Provincial Key Laboratory of Preventive Veterinary Medicine , Hangzhou, Zhejiang, China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-6787-0794","authenticated-orcid":true,"given":"Min","family":"Yue","sequence":"additional","affiliation":[{"name":"Department of Veterinary Medicine, Institute of Preventive Veterinary Sciences, Zhejiang University College of Animal Sciences , Hangzhou, Zhejiang, China"},{"name":"Hainan Institute of Zhejiang University , Sanya, Hainan, China"},{"name":"Zhejiang Provincial Key Laboratory of Preventive Veterinary Medicine , Hangzhou, Zhejiang, China"},{"name":"State Key Laboratory for Diagnosis and Treatment of Infectious Diseases, National Clinical Research Center for Infectious Diseases, National Medical Center for Infectious Diseases, The First Affiliated Hospital, Zhejiang University School of Medicine , Hangzhou, Zhejiang, China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-4813-5783","authenticated-orcid":false,"given":"Yan","family":"Li","sequence":"additional","affiliation":[{"name":"Department of Veterinary Medicine, Institute of Preventive Veterinary Sciences, Zhejiang University College of Animal Sciences , Hangzhou, Zhejiang, China"},{"name":"Hainan Institute of Zhejiang University , Sanya, Hainan, China"},{"name":"Zhejiang Provincial Key Laboratory of Preventive Veterinary Medicine , Hangzhou, Zhejiang, China"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"235","reference":[{"key":"e_1_3_2_2_2","first-page":"353","article-title":"\u00dcber eine neue Infektionskrankheit bei Haustieren","volume":"32","author":"Aujeszky A","year":"1902","unstructured":"Aujeszky A . 1902. \u00dcber eine neue Infektionskrankheit bei Haustieren. Zent bl Bakteriol Parasitenkd Infekt krankh 32:353\u2013357.","journal-title":"Zent bl Bakteriol Parasitenkd Infekt krankh"},{"key":"e_1_3_2_3_2","doi-asserted-by":"publisher","DOI":"10.1007\/s12250-020-00340-0"},{"key":"e_1_3_2_4_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.vetmic.2016.11.019"},{"key":"e_1_3_2_5_2","doi-asserted-by":"publisher","DOI":"10.1128\/JVI.52.2.583-590.1984"},{"key":"e_1_3_2_6_2","doi-asserted-by":"publisher","DOI":"10.1128\/MMBR.69.3.462-500.2005"},{"key":"e_1_3_2_7_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.mcp.2020.101605"},{"key":"e_1_3_2_8_2","doi-asserted-by":"publisher","DOI":"10.3201\/eid1911.130177"},{"key":"e_1_3_2_9_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.vaccine.2014.04.035"},{"key":"e_1_3_2_10_2","doi-asserted-by":"publisher","DOI":"10.3390\/v13040691"},{"key":"e_1_3_2_11_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.vetmic.2015.12.008"},{"key":"e_1_3_2_12_2","first-page":"42","article-title":"Experimental reduction of virulence of Aujeszky\u2019s disease virus","volume":"16","author":"Bartha A.","year":"1961","unstructured":"Bartha A. 1961. Experimental reduction of virulence of Aujeszky\u2019s disease virus. 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The safety, stability and Immunogenicity of double gene-negative mutant of pseudorabies virus strain (PRV HB-98). Chin J Vet Sci 26:165\u2013168.","journal-title":"Chin J Vet Sci"},{"key":"e_1_3_2_28_2","doi-asserted-by":"publisher","DOI":"10.1186\/s12917-016-0897-z"},{"key":"e_1_3_2_29_2","first-page":"1","article-title":"Immunization against Aujeszky\u2019s disease with live vaccine. I. Attenuation of virus and some properties of attenuated strains","volume":"8","author":"Skoda R","year":"1964","unstructured":"Skoda R , Brauner I , Sadecky E , Mayer V . 1964. Immunization against Aujeszky\u2019s disease with live vaccine. I. Attenuation of virus and some properties of attenuated strains. 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