{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,13]],"date-time":"2026-10-13T22:28:06Z","timestamp":1791930486273,"version":"4.3.4"},"reference-count":31,"publisher":"Walter de Gruyter GmbH","issue":"9","content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":[],"published-print":{"date-parts":[[2026,8,24]]},"abstract":"<jats:title>Abstract<\/jats:title>\n                  <jats:sec id=\"j_cclm-2025-1137_abs_001\">\n                    <jats:title>Objectives<\/jats:title>\n                    <jats:p>This study developed a pseudoviral human rhinovirus (HRV) reference material using pseudovirus technology and integrating RT-dPCR with HPLC-IDMS to achieve accurate HRV RNA quantification, aiming to standardize HRV detection across labs and platforms.<\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec id=\"j_cclm-2025-1137_abs_002\">\n                    <jats:title>Methods<\/jats:title>\n                    <jats:p>\n                      We developed an RT-digital PCR (RT-dPCR) reference method for HRV, incorporating reverse transcription efficiency (RTE) correction using HPLC-isotope dilution mass spectrometry (HPLC-IDMS), which enables value assignment of the well-characterized pseudoviral reference material (RM) containing the conserved\n                      <jats:italic>5\u2032UTR<\/jats:italic>\n                      \u00a0gene.\n                    <\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec id=\"j_cclm-2025-1137_abs_003\">\n                    <jats:title>Results<\/jats:title>\n                    <jats:p>\n                      The developed RT-dPCR method demonstrated high analytical sensitivity with a limit of detection (LoD) of eight copies\/reaction and a limit of quantification (LoQ) of 11 copies\/reaction. RTE was accurately determined as 101.25\u202f% using HPLC-IDMS, enabling metrologically traceable RNA quantification. The pseudoviral RM was characterized using the traceable RT-dPCR method; its reference value and expanded uncertainty were determined to be (3.12\u00a0\u00b1\u00a00.69)\u00a0\u00d7\u00a010\n                      <jats:sup>3<\/jats:sup>\n                      copies\/\u03bcL (k=2). The RM exhibited sufficient homogeneity and stability for 12\u00a0months at\u00a0\u221280\u202f\u00b0C, and its non-infectious nature coupled with its ability to simulate the entire workflow including nucleic acid extraction offers significant advantages.\n                    <\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec id=\"j_cclm-2025-1137_abs_004\">\n                    <jats:title>Conclusions<\/jats:title>\n                    <jats:p>This study provided essential tools for standardizing HRV detection across different laboratories and platforms. This work enables traceable RNA quantification in SI units and simulates the full analytical process, thereby enhancing the accuracy and comparability of results. Furthermore, this approach serves as a transferable model for developing reference materials and traceable quantification methods for other respiratory pathogens, contributing to advancements in molecular diagnostics and public health.<\/jats:p>\n                  <\/jats:sec>","DOI":"10.1515\/cclm-2025-1137","type":"journal-article","created":{"date-parts":[[2026,5,6]],"date-time":"2026-05-06T12:03:21Z","timestamp":1778069001000},"page":"1957-1965","source":"Crossref","is-referenced-by-count":1,"title":["Pseudovirus reference material and RT-digital PCR reference method for the quantification of human rhinovirus RNA with metrological traceability"],"prefix":"10.1515","volume":"64","author":[{"given":"Huijie","family":"Li","sequence":"first","affiliation":[{"name":"Center for Advanced Measurement of Science , National Institute of Metrology , Beijing , China"},{"name":"Shenzhen Institute for Technology Innovation , National Institute of Metrology , Shenzhen , 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National Institute of Metrology , Beijing , China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Shangjuan","family":"Wang","sequence":"additional","affiliation":[{"name":"Nanjing Institute of Measurement and Testing Technology , Nanjing , China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Lingxiang","family":"Zhu","sequence":"additional","affiliation":[{"name":"TargetingOne Corporation , Beijing , China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yi","family":"Yang","sequence":"additional","affiliation":[{"name":"Center for Advanced Measurement of Science , National Institute of Metrology , Beijing , China"},{"name":"Shenzhen Institute for Technology Innovation , National Institute of Metrology , Shenzhen , 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Beijing Academy of Science and Technology (Beijing Center for Physical & Chemical Analysis) , Beijing , China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Xun","family":"Chen","sequence":"additional","affiliation":[{"name":"School of Medicine Qinghua Yuan , Tsinghua University , Beijing , China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Haofeng","family":"Chen","sequence":"additional","affiliation":[{"name":"National Research Institute for Family Planning , Beijing , China"},{"name":"National Human Genetic Resources Center , Beijing , China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yong","family":"Guo","sequence":"additional","affiliation":[{"name":"School of Medicine Qinghua Yuan , Tsinghua University , Beijing , China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Huafang","family":"Gao","sequence":"additional","affiliation":[{"name":"National Research Institute for Family Planning , Beijing , 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