'Understanding the use of real time reverse trascriptase polymerase chain reaction (Rrt-PCR) for COVID 19 Diagnosis2
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Date
2020-08-01
Authors
Olalekan, A
Iwalokun, B
Adekunle, O
Makun, H
Mirabeau, T
Akinloye, O
Journal Title
Journal ISSN
Volume Title
Publisher
Pan African Journal of Life Sciences
Abstract
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
Description
Keywords
COVID -19, rRT-PCR, SARS-CoV-2, biological samples, target genes