{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,25]],"date-time":"2026-09-25T23:19:23Z","timestamp":1790378363422,"version":"4.1.0"},"reference-count":24,"publisher":"Frontiers Media SA","license":[{"start":{"date-parts":[[2026,6,4]],"date-time":"2026-06-04T00:00:00Z","timestamp":1780531200000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"content-domain":{"domain":["frontiersin.org"],"crossmark-restriction":true},"short-container-title":["Front. Cell. Infect. Microbiol."],"abstract":"<jats:sec>\n                    <jats:title>Background<\/jats:title>\n                    <jats:p>Northeastern China is a recognized hotspot for tick-borne viral diseases. However, there remains a lack of methods capable of simultaneously detecting these emerging and re-emerging tick-borne viruses.<\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Methods<\/jats:title>\n                    <jats:p>Primer and probe sets were designed using Beacon Designer 8.0, targeting conserved regions of viral genomes from Alongshan virus (ALSV), Tick-borne encephalitis virus (TBEV), Severe fever with thrombocytopenia syndrome virus (SFTSV), Beiji nairovirus (BJNV), Yezo virus (YEZV), and Songling virus (SGLV). Recombinant plasmids were constructed to assess assay sensitivity, while virus-positive cDNA from tick samples was used to verify specificity. The performance of these assays was further validated using field-collected tick samples, with results compared against established reference methods serving as gold standards.<\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Results<\/jats:title>\n                    <jats:p>Specific primer and probe sets were designed with amplicon lengths ranging from 83 to 199 bp. Optimization of the reaction components yielded final primer volumes of 0.2\u20130.5 \u03bcL and probe volumes of 0.4\u20131.0 \u03bcL (from 10 \u03bcM working stocks). Sensitivity analysis demonstrated a limit of detection (LOD) as low as 10 copies\/\u03bcL for all six viruses, while specificity testing confirmed no cross-reactivity among the targets. In validation trials, the established TaqMan RT-qPCR assays showed complete concordance with reference methods for ALSV (5\/15) and TBEV (4\/14). Notably, the TaqMan assays identified additional positive samples for SFTSV (n=1), BJNV (n=3), and YEZV (n=1) that were missed by the reference assays, particularly in samples with low viral loads. Conversely, two samples that tested positive for SGLV using SYBR Green RT-qPCR yielded negative results with the TaqMan RT-qPCR assay.<\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Conclusion<\/jats:title>\n                    <jats:p>We have successfully developed a suite of highly sensitive and specific single-plex TaqMan RT-qPCR assays for the rapid detection of six key tick-borne viruses in northeastern China. These assays facilitate efficient viral surveillance in tick populations and provide a diagnostic tool with significant potential for clinical applications.<\/jats:p>\n                  <\/jats:sec>","DOI":"10.3389\/fcimb.2026.1777543","type":"journal-article","created":{"date-parts":[[2026,6,4]],"date-time":"2026-06-04T05:41:06Z","timestamp":1780551666000},"update-policy":"https:\/\/doi.org\/10.3389\/crossmark-policy","source":"Crossref","is-referenced-by-count":2,"title":["Development and evaluation of single-plex TaqMan real-time quantitative PCR assays for the detection of six tick-borne pathogenic viruses in northeastern China"],"prefix":"10.3389","volume":"16","author":[{"given":"Shiyao","family":"Xu","sequence":"first","affiliation":[{"name":"Department of Infectious Diseases, Center of Infectious Diseases and Pathogen Biology, State Key Laboratory of Zoonotic Diseases, the First Hospital of Jilin University","place":["Changchun, Jilin, China"]},{"name":"College of Life Science, Northeast Forestry University","place":["Harbin, Heilongjiang, China"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Zheng","family":"Gui","sequence":"additional","affiliation":[{"name":"Department of Infectious Diseases, Center of Infectious Diseases and Pathogen Biology, State Key Laboratory of Zoonotic Diseases, the First Hospital of Jilin University","place":["Changchun, Jilin, China"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Ziyan","family":"Liu","sequence":"additional","affiliation":[{"name":"Department of Infectious Diseases, Center of Infectious Diseases and Pathogen Biology, State Key Laboratory of Zoonotic Diseases, the First Hospital of Jilin University","place":["Changchun, Jilin, China"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Kai","family":"Wang","sequence":"additional","affiliation":[{"name":"State Key Laboratory of Pathogen and Biosecurity, Changchun Veterinary Research Institute, Chinese Academy of Agricultural Sciences","place":["Changchun, Jilin, China"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Ning","family":"Liu","sequence":"additional","affiliation":[{"name":"Department of Infectious Diseases, Center of Infectious Diseases and Pathogen Biology, State Key Laboratory of Zoonotic Diseases, the First Hospital of Jilin University","place":["Changchun, Jilin, China"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yunzhi","family":"Peng","sequence":"additional","affiliation":[{"name":"Department of Infectious Diseases, Center of Infectious Diseases and Pathogen Biology, State Key Laboratory of Zoonotic Diseases, the First Hospital of Jilin University","place":["Changchun, Jilin, China"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Liang","family":"Li","sequence":"additional","affiliation":[{"name":"State Key Laboratory of Pathogen and Biosecurity, Changchun Veterinary Research Institute, Chinese Academy of Agricultural Sciences","place":["Changchun, Jilin, China"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Zedong","family":"Wang","sequence":"additional","affiliation":[{"name":"Department of Infectious Diseases, Center of Infectious Diseases and Pathogen Biology, State Key Laboratory of Zoonotic Diseases, the First Hospital of Jilin University","place":["Changchun, Jilin, China"]},{"name":"International Center of Future Science, Jilin University","place":["Changchun, Jilin, China"]}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1965","published-online":{"date-parts":[[2026,6,4]]},"reference":[{"key":"B1","doi-asserted-by":"publisher","DOI":"10.1080\/22221751.2024.2404271","article-title":"Epidemiology and global spread of emerging tick-borne Alongshan virus","volume":"13","author":"Gomer","year":"2024","journal-title":"Emerg. 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