{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,8,18]],"date-time":"2026-08-18T00:42:11Z","timestamp":1787013731311,"version":"build-2736575974"},"reference-count":52,"publisher":"Wiley","issue":"7","license":[{"start":{"date-parts":[[2022,3,22]],"date-time":"2022-03-22T00:00:00Z","timestamp":1647907200000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/100010661","name":"Horizon 2020 Framework Programme","doi-asserted-by":"publisher","award":["871029"],"award-info":[{"award-number":["871029"]}],"id":[{"id":"10.13039\/100010661","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100005357","name":"Agent\u00fara na Podporu V\u00fdskumu a V\u00fdvoja","doi-asserted-by":"publisher","award":["PP\u2010COVID\u201020\u20100017"],"award-info":[{"award-number":["PP\u2010COVID\u201020\u20100017"]}],"id":[{"id":"10.13039\/501100005357","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100005357","name":"Agent\u00fara na Podporu V\u00fdskumu a V\u00fdvoja","doi-asserted-by":"publisher","award":["PP\u2010COVID\u201020\u20100116"],"award-info":[{"award-number":["PP\u2010COVID\u201020\u20100116"]}],"id":[{"id":"10.13039\/501100005357","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":["enviromicro-journals.onlinelibrary.wiley.com"],"crossmark-restriction":true},"short-container-title":["Microbial Biotechnology"],"published-print":{"date-parts":[[2022,7]]},"abstract":"<jats:title>Summary<\/jats:title>\n                  <jats:p>Sensitive and accurate RT\u2010qPCR tests are the primary diagnostic tools to identify SARS\u2010CoV\u20102\u2010infected patients. While many SARS\u2010CoV\u20102 RT\u2010qPCR tests are available, there are significant differences in test sensitivity, workflow (e.g. hands\u2010on\u2010time), gene targets and other functionalities that users must consider. Several publicly available protocols shared by reference labs and public health authorities provide useful tools for SARS\u2010CoV\u20102 diagnosis, but many have shortcomings related to sensitivity and laborious workflows. Here, we describe a series of SARS\u2010CoV\u20102 RT\u2010qPCR tests that are originally based on the protocol targeting regions of the RNA\u2010dependent RNA polymerase (RdRp) and envelope (E) coding genes developed by the Charit\u00e9 Berlin. We redesigned the primers\/probes, utilized locked nucleic acid nucleotides, incorporated dual probe technology and conducted extensive optimizations of reaction conditions to enhance the sensitivity and specificity of these tests. By incorporating an RNase P internal control and developing multiplexed assays for distinguishing SARS\u2010CoV\u20102 and influenza A and B, we streamlined the workflow to provide quicker results and reduced consumable costs. Some of these tests use modified enzymes enabling the formulation of a room temperature\u2010stable master mix and lyophilized positive control, thus increasing the functionality of the test and eliminating cold chain shipping and storage. Moreover, a rapid, RNA extraction\u2010free version enables high sensitivity detection of SARS\u2010CoV\u20102 in about an hour using minimally invasive, self\u2010collected gargle samples. These RT\u2010qPCR assays can easily be implemented in any diagnostic laboratory and can provide a powerful tool to detect SARS\u2010CoV\u20102 and the most common seasonal influenzas during the vaccination phase of the pandemic.<\/jats:p>","DOI":"10.1111\/1751-7915.14031","type":"journal-article","created":{"date-parts":[[2022,3,22]],"date-time":"2022-03-22T12:04:40Z","timestamp":1647950680000},"page":"1995-2021","update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":6,"title":["Sequential development of several RT\u2010qPCR tests using LNA nucleotides and dual probe technology to differentiate SARS\u2010CoV\u20102 from influenza A and B"],"prefix":"10.1111","volume":"15","author":[{"ORCID":"https:\/\/orcid.org\/0000-0001-5904-2377","authenticated-orcid":false,"given":"Monika","family":"Radv\u00e1nszka","sequence":"first","affiliation":[{"name":"MultiplexDX, s.r.o. 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