{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,8,10]],"date-time":"2026-08-10T09:28:35Z","timestamp":1786354115259,"version":"build-2736575974"},"reference-count":29,"publisher":"MDPI AG","issue":"8","license":[{"start":{"date-parts":[[2026,8,8]],"date-time":"2026-08-08T00:00:00Z","timestamp":1786147200000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"the Scientific Research Project of Anhui Academy of Agricultural Sciences","award":["No.2026YL037"],"award-info":[{"award-number":["No.2026YL037"]}]},{"award":["No.2026YL037"],"award-info":[{"award-number":["No.2026YL037"]}],"id":[{"id":"https:\/\/ror.org\/01pw5qp76","id-type":"ROR","asserted-by":"publisher"}]},{"name":"the China Agriculture Research System of MOF and MARA","award":["No. CARS-39"],"award-info":[{"award-number":["No. CARS-39"]}]},{"award":["No. CARS-39"],"award-info":[{"award-number":["No. CARS-39"]}],"id":[{"id":"https:\/\/ror.org\/03p5eky76","id-type":"ROR","asserted-by":"publisher"}]},{"name":"Chuzhou, Multidisciplinary, Observation and Research Station, Ministry of Agriculture and Rural Affairs","award":["No. MARA-ORS046-AH007"],"award-info":[{"award-number":["No. MARA-ORS046-AH007"]}]},{"name":"Chuzhou, Multidisciplinary, Observation and Research Station, Ministry of Agriculture and Rural Affairs","award":["2026YL084"],"award-info":[{"award-number":["2026YL084"]}]},{"award":["No. MARA-ORS046-AH007"],"award-info":[{"award-number":["No. MARA-ORS046-AH007"]}],"id":[{"id":"https:\/\/ror.org\/05ckt8b96","id-type":"ROR","asserted-by":"publisher"}]},{"award":["2026YL084"],"award-info":[{"award-number":["2026YL084"]}],"id":[{"id":"https:\/\/ror.org\/05ckt8b96","id-type":"ROR","asserted-by":"publisher"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Biosensors"],"abstract":"<jats:p>Goose astrovirus genotype 2 (GAstV-2) is an important pathogen associated with gosling gout, and rapid detection is useful for early diagnosis and field surveillance. In this study, a visual assay for GAstV-2 detection was developed by combining one-step reverse transcription multienzyme isothermal rapid amplification (MIRA) with the nucleic acid cleavage activity of Pyrococcus furiosus Argonaute (PfAgo). MIRA primers and specific guide DNAs were designed based on a conserved region of the GAstV-2 ORF1b gene, and the PfAgo reaction conditions were optimized. The optimal reaction contained 1.0 \u03bcM gDNA, 0.6 \u03bcM PfAgo, and 1.0 mM MnCl2. Using recombinant pUC57-ORF1b plasmid DNA as the template, the lowest detectable plasmid concentration under the tested conditions was 1.0 \u00d7 100 copies\/\u03bcL. In the specificity assay, only GAstV-2 produced a positive signal, with no cross-reaction observed with GAstV-1, Tembusu virus, H9-subtype avian influenza virus, goose circovirus, fowl adenovirus serotype 4, or goose parvovirus. The assay was further tested with 23 clinical samples suspected of GAstV-2 infection. In a preliminary evaluation of 23 clinical samples, the RT-MIRA-PfAgo results were concordant with those obtained by conventional RT-PCR and RT-qPCR. Overall, the RT-MIRA-PfAgo assay provided sensitive and specific GAstV detection within a short time, without requiring programmed thermal cycling or an expensive real-time PCR instrument for routine endpoint detection. This method may be useful for GAstV-2 detection in basic laboratories and field settings.<\/jats:p>","DOI":"10.3390\/bios16080433","type":"journal-article","created":{"date-parts":[[2026,8,10]],"date-time":"2026-08-10T08:08:53Z","timestamp":1786349333000},"page":"433","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":0,"title":["Development of a Visual Rapid Assay for Novel Goose Astrovirus Detection Based on RT-MIRA -PfAgo"],"prefix":"10.3390","volume":"16","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-1163-1305","authenticated-orcid":false,"given":"Dongdong","family":"Yin","sequence":"first","affiliation":[{"name":"Anhui Provincial Key Laboratory of Livestock and Poultry Product Safety, Institute of Animal Husbandry and Veterinary Science, Anhui Academy of Agricultural Sciences, Livestock and Poultry Epidemic Diseases Research Center of Anhui Province, Hefei 230031, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Xinjun","family":"Chen","sequence":"additional","affiliation":[{"name":"Yebio Bioengineering Co., Ltd of Qingdao, Qingdao 266113, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Zhixing","family":"Cheng","sequence":"additional","affiliation":[{"name":"Yebio Bioengineering Co., Ltd of Qingdao, Qingdao 266113, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yu","family":"Liu","sequence":"additional","affiliation":[{"name":"Yebio Bioengineering Co., Ltd of Qingdao, Qingdao 266113, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yin","family":"Dai","sequence":"additional","affiliation":[{"name":"Anhui Provincial Key Laboratory of Livestock and Poultry Product Safety, Institute of Animal Husbandry and Veterinary Science, Anhui Academy of Agricultural Sciences, Livestock and Poultry Epidemic Diseases Research Center of Anhui Province, Hefei 230031, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Xuehuai","family":"Shen","sequence":"additional","affiliation":[{"name":"Anhui Provincial Key Laboratory of Livestock and Poultry Product Safety, Institute of Animal Husbandry and Veterinary Science, Anhui Academy of Agricultural Sciences, Livestock and Poultry Epidemic Diseases Research Center of Anhui Province, Hefei 230031, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Xiaocheng","family":"Pan","sequence":"additional","affiliation":[{"name":"Anhui Provincial Key Laboratory of Livestock and Poultry Product Safety, Institute of Animal Husbandry and Veterinary Science, Anhui Academy of Agricultural Sciences, Livestock and Poultry Epidemic Diseases Research Center of Anhui Province, Hefei 230031, China"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2026,8,8]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"53","DOI":"10.1016\/j.vetmic.2018.05.006","article-title":"Novel Goose Astrovirus Associated Gout in Gosling, China","volume":"220","author":"Niu","year":"2018","journal-title":"Vet. 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