{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,7,1]],"date-time":"2026-07-01T08:39:45Z","timestamp":1782895185330,"version":"3.54.5"},"reference-count":25,"publisher":"Wiley","issue":"1","license":[{"start":{"date-parts":[[2026,7,1]],"date-time":"2026-07-01T00:00:00Z","timestamp":1782864000000},"content-version":"vor","delay-in-days":181,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"},{"start":{"date-parts":[[2026,1,1]],"date-time":"2026-01-01T00:00:00Z","timestamp":1767225600000},"content-version":"tdm","delay-in-days":0,"URL":"http:\/\/doi.wiley.com\/10.1002\/tdm_license_1.1"}],"funder":[{"DOI":"10.13039\/501100004704","name":"National Research Council of Thailand","doi-asserted-by":"publisher","award":["N41A640189"],"award-info":[{"award-number":["N41A640189"]}],"id":[{"id":"10.13039\/501100004704","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100002873","name":"Chulalongkorn University","doi-asserted-by":"publisher","award":["HEA_FF_69_052_3100_004"],"award-info":[{"award-number":["HEA_FF_69_052_3100_004"]}],"id":[{"id":"10.13039\/501100002873","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":["onlinelibrary.wiley.com"],"crossmark-restriction":true},"short-container-title":["Transboundary and Emerging Diseases"],"published-print":{"date-parts":[[2026,1]]},"abstract":"<jats:p>\n                    Feline bocaviruses (FBoVs) are increasingly recognized for their global distribution and potential importance in feline health. Three distinct species\u2014FBoV\u20101, FBoV\u20102, and FBoV\u20103\u2014have been identified; however, the lack of efficient molecular assays has limited the ability to clarify their species distribution, prevalence, and viral loads. In this study, we developed a multiplex quantitative PCR (qPCR) assay targeting the\n                    <jats:italic>NS1<\/jats:italic>\n                    gene for the simultaneous detection, differentiation, and quantification of all three FBoV species. The assay demonstrated high analytical sensitivity, with limits of detection of 1.6\u2009\u00d7\u200910\n                    <jats:sup>3<\/jats:sup>\n                    \u2009copies\/\u03bcL for FBoV\u20101 and 1.6\u2009\u00d7\u200910\n                    <jats:sup>2<\/jats:sup>\n                    \u2009copies\/\u03bcL for both FBoV\u20102 and FBoV\u20103. No cross\u2010reactivity was observed with other feline or non\u2010feline viruses, and the assay showed strong repeatability and reproducibility. Diagnostic evaluation using 135 archived feline oropharyngeal and fecal samples yielded a sensitivity of 95.2% and a specificity of 100% compared with previously established singleplex qPCR assays. Importantly, the multiplex format reduced time, cost, and labor while maintaining high diagnostic accuracy. This study presents the first multiplex qPCR assay capable of simultaneously detecting, differentiating, and quantifying all three FBoV species, providing a rapid, sensitive, and specific tool for routine diagnostics and large\u2010scale epidemiological investigations.\n                  <\/jats:p>","DOI":"10.1155\/tbed\/3455556","type":"journal-article","created":{"date-parts":[[2026,7,1]],"date-time":"2026-07-01T07:27:45Z","timestamp":1782890865000},"update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":0,"title":["The Multiplex qPCR Assay Enables Simultaneous Detection, Differentiation, and Quantification of Feline Bocavirus\u20101, \u20102, and \u20103 in Clinical Samples"],"prefix":"10.1155","volume":"2026","author":[{"ORCID":"https:\/\/orcid.org\/0000-0003-3236-1399","authenticated-orcid":false,"given":"Pattiya","family":"Lohavicharn","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-1895-8992","authenticated-orcid":false,"given":"Tanit","family":"Kasantikul","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-5148-4696","authenticated-orcid":false,"given":"Chutchai","family":"Piewbang","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-4888-2677","authenticated-orcid":false,"given":"Somporn","family":"Techangamsuwan","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"311","published-online":{"date-parts":[[2026,7,1]]},"reference":[{"key":"e_1_2_10_1_2","doi-asserted-by":"publisher","DOI":"10.1093\/nar\/gkx932"},{"key":"e_1_2_10_2_2","doi-asserted-by":"publisher","DOI":"10.1007\/s00705-013-1914-1"},{"key":"e_1_2_10_3_2","doi-asserted-by":"publisher","DOI":"10.1186\/s12985-018-1034-3"},{"key":"e_1_2_10_4_2","doi-asserted-by":"publisher","DOI":"10.1099\/vir.0.042531-0"},{"key":"e_1_2_10_5_2","doi-asserted-by":"publisher","DOI":"10.1007\/s00705-016-2972-y"},{"key":"e_1_2_10_6_2","doi-asserted-by":"publisher","DOI":"10.1186\/1743-422X-10-54"},{"key":"e_1_2_10_7_2","doi-asserted-by":"publisher","DOI":"10.1038\/s41598-019-52902-2"},{"key":"e_1_2_10_8_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.vetmic.2014.04.005"},{"key":"e_1_2_10_9_2","doi-asserted-by":"publisher","DOI":"10.1111\/tbed.14646"},{"key":"e_1_2_10_10_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.virol.2024.110188"},{"key":"e_1_2_10_11_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.cimid.2021.101676"},{"key":"e_1_2_10_12_2","doi-asserted-by":"crossref","unstructured":"WangY. LiW. andGuoX. et al.Development of SYBR Green I-Based Polymerase Chain Reaction for Feline Bocavirus 1 Detection 2021 11 no. 2 https:\/\/doi.org\/10.1007\/s13205-020-02577-8 61.","DOI":"10.1007\/s13205-020-02577-8"},{"key":"e_1_2_10_13_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.mcp.2020.101647"},{"key":"e_1_2_10_14_2","doi-asserted-by":"crossref","unstructured":"WangY. PanY. andWuJ. et al.Simultaneous Detection of Feline Parvovirus and Feline Bocavirus Using SYBR Green I-Based Duplex Real-Time Polymerase Chain Reaction 2021 11 no. 9 https:\/\/doi.org\/10.1007\/s13205-021-02947-w 400.","DOI":"10.1007\/s13205-021-02947-w"},{"key":"e_1_2_10_15_2","doi-asserted-by":"publisher","DOI":"10.3389\/fvets.2022.1005759"},{"key":"e_1_2_10_16_2","doi-asserted-by":"publisher","DOI":"10.1093\/bib\/bbx108"},{"key":"e_1_2_10_17_2","doi-asserted-by":"crossref","first-page":"31","DOI":"10.1007\/978-1-4939-2365-6_3","volume-title":"PCR Primer Design","author":"Rodr\u00edguez A.","year":"2015"},{"key":"e_1_2_10_18_2","unstructured":"BLASTn Program https:\/\/blast.ncbi.nlm.nih.gov."},{"key":"e_1_2_10_19_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.meegid.2024.105675"},{"key":"e_1_2_10_20_2","doi-asserted-by":"publisher","DOI":"10.1177\/1098612X251384767"},{"key":"e_1_2_10_21_2","doi-asserted-by":"publisher","DOI":"10.1111\/tbed.14664"},{"key":"e_1_2_10_22_2","unstructured":"Random Number Generator https:\/\/www.calculatorsoup.com\/calculators\/statistics\/random-number-generator.php."},{"key":"e_1_2_10_23_2","doi-asserted-by":"publisher","DOI":"10.1371\/journal.pone.0255425"},{"key":"e_1_2_10_24_2","doi-asserted-by":"publisher","DOI":"10.1093\/infdis\/jiaf090"},{"key":"e_1_2_10_25_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.mimet.2020.106051"}],"container-title":["Transboundary and Emerging Diseases"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/onlinelibrary.wiley.com\/doi\/pdf\/10.1155\/tbed\/3455556","content-type":"application\/pdf","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/onlinelibrary.wiley.com\/doi\/full-xml\/10.1155\/tbed\/3455556","content-type":"application\/xml","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/onlinelibrary.wiley.com\/doi\/pdf\/10.1155\/tbed\/3455556","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2026,7,1]],"date-time":"2026-07-01T07:27:56Z","timestamp":1782890876000},"score":1,"resource":{"primary":{"URL":"https:\/\/onlinelibrary.wiley.com\/doi\/10.1155\/tbed\/3455556"}},"subtitle":[],"editor":[{"given":"Nanhua","family":"Chen","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"editor"}]}],"short-title":[],"issued":{"date-parts":[[2026,1]]},"references-count":25,"journal-issue":{"issue":"1","published-print":{"date-parts":[[2026,1]]}},"alternative-id":["10.1155\/tbed\/3455556"],"URL":"https:\/\/doi.org\/10.1155\/tbed\/3455556","archive":["Portico"],"relation":{},"ISSN":["1865-1674","1865-1682"],"issn-type":[{"value":"1865-1674","type":"print"},{"value":"1865-1682","type":"electronic"}],"subject":[],"published":{"date-parts":[[2026,1]]},"assertion":[{"value":"2026-03-31","order":0,"name":"received","label":"Received","group":{"name":"publication_history","label":"Publication History"}},{"value":"2026-06-10","order":2,"name":"accepted","label":"Accepted","group":{"name":"publication_history","label":"Publication History"}},{"value":"2026-07-01","order":3,"name":"published","label":"Published","group":{"name":"publication_history","label":"Publication History"}}],"article-number":"3455556"}}