Real-Time Polymerase Chain Reaction: Current Techniques, Applications, and Role in COVID-19 Diagnosis

Genes (Basel). 2022 Dec 16;13(12):2387. doi: 10.3390/genes13122387.

Abstract

Successful detection of the first SARS-CoV-2 cases using the real-time polymerase chain reaction (real-time PCR) method reflects the power and usefulness of this technique. Real-time PCR is a variation of the PCR assay to allow monitoring of the PCR progress in actual time. PCR itself is a molecular process used to enzymatically synthesize copies in multiple amounts of a selected DNA region for various purposes. Real-time PCR is currently one of the most powerful molecular approaches and is widely used in biological sciences and medicine because it is quantitative, accurate, sensitive, and rapid. Current applications of real-time PCR include gene expression analysis, mutation detection, detection and quantification of pathogens, detection of genetically modified organisms, detection of allergens, monitoring of microbial degradation, species identification, and determination of parasite fitness. The technique has been used as a gold standard for COVID-19 diagnosis. Modifications of the standard real-time PCR methods have also been developed for particular applications. This review aims to provide an overview of the current applications of the real-time PCR technique, including its role in detecting emerging viruses such as SARS-CoV-2.

Keywords: COVID-19; SARS-CoV-2; molecular diagnosis; polymerase chain reaction; quantitative PCR; real-time PCR.

Publication types

  • Review

MeSH terms

  • COVID-19 Testing
  • COVID-19* / diagnosis
  • COVID-19* / genetics
  • Humans
  • RNA, Viral / genetics
  • Real-Time Polymerase Chain Reaction / methods
  • SARS-CoV-2 / genetics

Substances

  • RNA, Viral

Grants and funding

This research did not receive any specific grant from funding agencies in the public, commercial, or not-for-profit sectors.