Background: Adequate knowledge of real time Reverse Transcriptase-Polymerase Chain Re-action (rRT-PCR) is critical for accurate implementation of the assay, interpretation of results and report-ing. 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 narrative review was carried out to describe the principles of rRT-PCR, provide an update on the landscape of rRT-PCR protocols and elucidate the process control involved in pre-analytical, analytical and post-analytical stages of COVID-19 testing .
Review Findings: The rRT-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 proto-cols.
Conclusion: rRT-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 rRT-PCR protocol prior to deployment for COVID-19 testing and research into the development of alternative testing protocols are strongly recommended
The Development and Application of a Custom Robotic Biomechanical Testing Platform Employing Real-time Load-control to Compare Spinal Biomechanical Testing Protocols: Pure Moment, Ideal Follower Load, and a Novel Trunk Weight Protocol