{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,15]],"date-time":"2026-09-15T05:02:27Z","timestamp":1789448547467,"version":"build-2803163510"},"reference-count":25,"publisher":"MDPI AG","issue":"9","license":[{"start":{"date-parts":[[2026,9,12]],"date-time":"2026-09-12T00:00:00Z","timestamp":1789171200000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/501100018806","name":"Scientific and Technological Project on Laboratory Animals of Hubei Province","doi-asserted-by":"publisher","award":["2024CFC002"],"award-info":[{"award-number":["2024CFC002"]}],"id":[{"id":"10.13039\/501100018806","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100012166","name":"National Key Research and Development Program","doi-asserted-by":"crossref","award":["2021YFF0702002"],"award-info":[{"award-number":["2021YFF0702002"]}],"id":[{"id":"10.13039\/501100012166","id-type":"DOI","asserted-by":"crossref"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Vet. Sci.","Veterinary Sciences"],"abstract":"<jats:p>Macacine alphaherpesvirus 1 (McHV-1, commonly known as Monkey B Virus) is a fatal zoonotic pathogen naturally prevalent in macaques. In this study, we developed and conducted a proof-of-concept analytical validation of a HotStart Loop-Mediated Isothermal Amplification (LAMP) assay targeting a conserved, moderately GC-rich (61\u201362%) region of the UL1 gene in McHV-1. The reaction was optimized at 70 \u00b0C using engineered HotStart Bst 3.2\/4.2 DNA Polymerase, tracked via real-time fluorescence and closed-tube visual colorimetric detection (pre-added L-HNB dye) without post-amplification tube opening to reduce the risk of post-amplification aerosol carry-over contamination. The assay achieved a preliminary practical analytical limit of detection (analytical LOD) of 100 copies\/reaction (3\/3 replicates) within a 15\u201330 min diagnostic cutoff window, matching the practical sensitivity of reference qPCR. Strict analytical specificity was demonstrated against a comprehensive panel of human herpesviruses (HHV-1 through HHV-5) and host macaque genomic DNA. As a single-target analytical prototype, this visual HotStart LAMP platform provides a rapid, equipment-free molecular tool with promising potential for McHV-1 surveillance in resource-limited settings and macaque breeding facilities upon further clinical validation.<\/jats:p>","DOI":"10.3390\/vetsci13090951","type":"journal-article","created":{"date-parts":[[2026,9,14]],"date-time":"2026-09-14T13:05:26Z","timestamp":1789391126000},"page":"951","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":0,"title":["Development and Proof-of-Concept Analytical Validation of a HotStart Loop-Mediated Isothermal Amplification (LAMP) Assay Targeting the UL1 Gene for Macacine alphaherpesvirus 1 (McHV-1)"],"prefix":"10.3390","volume":"13","author":[{"given":"Yang","family":"Xiao","sequence":"first","affiliation":[{"name":"Changde Vocational Technical College, Changde 415000, China"}],"role":[{"vocabulary":"crossref","role":"author"},{"role":"conceptualization","vocabulary":"credit"},{"role":"writing-original-draft","vocabulary":"credit"},{"role":"writing-review-editing","vocabulary":"credit"},{"role":"data-curation","vocabulary":"credit"},{"role":"formal-analysis","vocabulary":"credit"}]},{"given":"Lianxiang","family":"Guo","sequence":"additional","affiliation":[{"name":"Suzhou Xishan Biotech Inc., Suzhou 215021, China"}],"role":[{"vocabulary":"crossref","role":"author"},{"role":"conceptualization","vocabulary":"credit"},{"role":"writing-original-draft","vocabulary":"credit"},{"role":"writing-review-editing","vocabulary":"credit"},{"role":"data-curation","vocabulary":"credit"},{"role":"formal-analysis","vocabulary":"credit"},{"role":"resources","vocabulary":"credit"}]},{"given":"Jingyi","family":"Zhang","sequence":"additional","affiliation":[{"name":"Center for Animal Experiment\/ABSL-3 Laboratory, Wuhan University, Wuhan 430071, China"}],"role":[{"vocabulary":"crossref","role":"author"},{"role":"data-curation","vocabulary":"credit"}]},{"given":"Zhang","family":"Zhang","sequence":"additional","affiliation":[{"name":"Center for Animal Experiment\/ABSL-3 Laboratory, Wuhan University, Wuhan 430071, China"}],"role":[{"vocabulary":"crossref","role":"author"},{"role":"data-curation","vocabulary":"credit"}]},{"given":"Ziwen","family":"Long","sequence":"additional","affiliation":[{"name":"Center for Animal Experiment\/ABSL-3 Laboratory, Wuhan University, Wuhan 430071, China"}],"role":[{"vocabulary":"crossref","role":"author"},{"role":"data-curation","vocabulary":"credit"}]},{"given":"Hu","family":"Liu","sequence":"additional","affiliation":[{"name":"Center for Animal Experiment\/ABSL-3 Laboratory, Wuhan University, Wuhan 430071, China"}],"role":[{"vocabulary":"crossref","role":"author"},{"role":"data-curation","vocabulary":"credit"}]},{"given":"Yaoming","family":"Li","sequence":"additional","affiliation":[{"name":"Applied Biotechnology Research Center, Wuhan University of Bioengineering, Wuhan 430415, China"}],"role":[{"vocabulary":"crossref","role":"author"},{"role":"writing-review-editing","vocabulary":"credit"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-0877-9093","authenticated-orcid":false,"given":"Rong","family":"Bao","sequence":"additional","affiliation":[{"name":"Center for Animal Experiment\/ABSL-3 Laboratory, Wuhan University, Wuhan 430071, China"}],"role":[{"vocabulary":"crossref","role":"author"},{"role":"project-administration","vocabulary":"credit"},{"role":"supervision","vocabulary":"credit"},{"role":"funding-acquisition","vocabulary":"credit"},{"role":"writing-original-draft","vocabulary":"credit"},{"role":"writing-review-editing","vocabulary":"credit"},{"role":"formal-analysis","vocabulary":"credit"},{"role":"data-curation","vocabulary":"credit"},{"role":"conceptualization","vocabulary":"credit"},{"role":"investigation","vocabulary":"credit"}]}],"member":"1968","published-online":{"date-parts":[[2026,9,12]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"5248420","DOI":"10.1155\/2018\/5248420","article-title":"Questioning the extreme neurovirulence of monkey B virus (Macacine alphaherpesvirus 1)","volume":"2018","author":"Eberle","year":"2018","journal-title":"Adv. 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