{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,15]],"date-time":"2026-09-15T17:52:00Z","timestamp":1789494720598,"version":"build-2803163510"},"reference-count":29,"publisher":"Wiley","issue":"12","license":[{"start":{"date-parts":[[2020,2,14]],"date-time":"2020-02-14T00:00:00Z","timestamp":1581638400000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/onlinelibrary.wiley.com\/termsAndConditions#vor"}],"content-domain":{"domain":["onlinelibrary.wiley.com"],"crossmark-restriction":true},"short-container-title":["Journal of Medical Virology"],"published-print":{"date-parts":[[2020,12]]},"abstract":"<jats:title>Abstract<\/jats:title>\n                  <jats:p>Multiplex real\u2010time quantitative polymerase chain reaction (mRT\u2010qPCR) assay is commonly used to detect respiratory viruses, however, the sensitivity is limited for most reports. A panel of locked nucleic acid based multiplex closed one\u2010tube nested real\u2010time PCR (mOTNRT\u2010PCR) assay consisting of five separate internally controlled RT\u2010qPCR assays was developed for detection of 14 respiratory viruses. The sensitivity and reproducibility of mOTNRT\u2010PCR panel were evaluated using plasmid standards and the specificity was evaluated using clinical samples. The clinical performance of mOTNRT\u2010PCR panel was further evaluated with 468 samples collected from patients with an acute respiratory infection and compared with individual real\u2010time PCR (RT\u2010qPCR) assay. The analytical sensitivities of mOTNRT\u2010PCR panel ranged from 2 to 20 copies\/reaction, and no cross\u2010reaction with common respiratory viruses was observed. The coefficients of variation of intra\u2010assay and inter\u2010assay were between 0.35% and 8.29%. Totally 35 clinical samples detected by mOTNRT\u2010PCR assay panel were missed by RT\u2010qPCR and confirmed true positive by sequencing of nested PCR products. The mOTNRT\u2010PCR assay panel provides a more sensitive and high\u2010throughput method for the detection of 14 respiratory viruses.<\/jats:p>","DOI":"10.1002\/jmv.25686","type":"journal-article","created":{"date-parts":[[2020,1,25]],"date-time":"2020-01-25T04:21:26Z","timestamp":1579926086000},"page":"3073-3080","update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":8,"title":["Development and evaluation of a panel of multiplex one\u2010tube nested real time PCR assay for simultaneous detection of 14 respiratory viruses in five reactions"],"prefix":"10.1002","volume":"92","author":[{"given":"Li","family":"Zhao","sequence":"first","affiliation":[{"name":"Department of Blood Transfusion Children's Hospital of Hebei Province  Shijiazhuang Hebei China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Gui\u2010xia","family":"Li","sequence":"additional","affiliation":[{"name":"Institute of Pediatric Research Children's 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