PMC:7235801 / 7833-10711
Annnotations
{"target":"http://pubannotation.org/docs/sourcedb/PMC/sourceid/7235801","sourcedb":"PMC","sourceid":"7235801","source_url":"https://www.ncbi.nlm.nih.gov/pmc/7235801","text":"The most widely used IVD assays for the confirmatory diagnosis of COVID-19 are based on RT-PCR that have been employed globally to deal with the pandemic. A novel and robust real-time RT-PCR assay was developed by Tib-Molbiol, Germany, in collaboration with various partners by the 2nd week of January 2020 [14]. It was highly specific for SARS-CoV-2 RNA and did not cross-react with other coronaviruses. The test detects the SARS-CoV-2 RNA via envelope (E) and RNA-dependent RNA polymerase (RdRp) gene assays. The E-gene assay was used for first line screening, while the RdRp gene assay was employed for confirmatory testing. In another approach, the researchers developed one-step RT-PCR assays to detect ORF1b and N regions of SARS-CoV-2 in 1 h and 15 min [15]. The N gene assay was used for screening, while Orf1b was used as confirmatory test. However, as the assay involved the detection of ORF1b and N regions that are highly conserved among sarbecoviruses, it could also bind to SARS-CoV and other closely-related viruses. The authors mentioned that they could distinguish the SARS-CoV-2 from SARS-CoV via sequence analysis of positive amplicons if the RT-PCR results are positive. Of interest was the development of real-time RT-PCR assay for the detection of RdRp/helicase (H) genes of SARS-CoV-2, which did not cross-react with other human coronaviruses and respiratory viruses [16]. Due to its high sensitivity, it can be used for COVID-19 detection in samples that have low viral loads and for testing saliva, plasma, and upper respiratory tract samples. Many RT-PCR assays have already been developed by many companies and researchers worldwide. The most prominent real-time RT-PCR rapid test is the Xpert® Xpress SARS-CoV-2 test by Cepheid, USA, which provides results in just 45 min using GenXpert benchtop system [17]. It is a rapid and automated point-of-care (POC) molecular test that enables the qualitative detection of SARS-CoV-2 in nasopharyngeal swab, nasal wash, or aspirate specimens from suspects. The test requires only a minute for sample preparation, employs Cepheid’s Xpert® Xpress cartridge technology, and targets multiple regions of the viral genome. It has also received Food and Drug Administration (FDA) emergency use authorization (EUA). Another prominent advance is the Vivalytic COVID-19 test by Bosch, Germany [18], which detects SARS-CoV-2 infection in suspects in less than 2.5 h. The test was developed by Bosch in collaboration with Randox Laboratories, UK. It is a fully automated and rapid POC molecular test that can simultaneously detect SARS-CoV-2 and nine other respiratory viruses, including influenza A and B. The sample is taken from the nose or throat of suspect using a swab and placed inside a Vivalytic cartridge containing all the assay reagents. The cartridge is then plugged into a small benchtop Vivalytic 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