Department of Medical Laboratory Science

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Browsing Department of Medical Laboratory Science by Author "Adekunle, O"

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    Open Access
    'Understanding the use of real time reverse trascriptase polymerase chain reaction (Rrt-PCR) for COVID 19 Diagnosis2
    (Pan African Journal of Life Sciences, 2020-08-01) Olalekan, A; Iwalokun, B; Adekunle, O; Makun, H; Mirabeau, T; Akinloye, O
    Background: Adequ ate know ledge of r eal tim e Rever se Tr anscr iptase-Polymerase Chain Reaction (rRT-PCR) is critical for accurate implementation of the assay, interpretation of results and reporting. This mini-review describes the principles, procedures, and level of development of rRT-PCR assays for the control of the COVID-19 pandemic. Methods: A n ar r ative r eview w as car r ied out to descr ibe th e pr inciples of r RT-PCR, provide an update on the landscape of rRT-PCR protocols and elucidate the process control involved in preanalytical, analytical and post-analytical stages of COVID-19 testing . Review Findings: Th e r RT-PCR is currently considered to be the acceptable standard for confirming COVID-19 diagnosis based on SARS-CoV-2 RNA detection via conversion to cDNA and amplification of target genes in real time using sequence specific TaqMan® probes. Available evidence indicates that different rRT-PCR protocols varying in number and type of target genes within SARS-CoV-2 genome are currently available for validation and emergency use approval (EUA) in pandemic countries. A total of 1 – 3 target genes, comprising the ORF1a, ORF1b, RNA dependent RNA polymerase (RdRp), Nucleoplasid protein gene (N), Spike glycoprotein gene (S) and Envelope protein gene (E) are detected by these protocols. Conclusion: r RT-PCR remains the most sensitive method for confirming, monitoring and managing COVID-19 disease in the ongoing pandemic in all affected countries. The need for validation of every rRTPCR protocol prior to deployment for COVID-19 testing and research into the development of alternative testing protocols are strongly recommended