A BRIEF OVERVIEW OF COVID-19 DIAGNOSIS BY REAL-TIME RT-PCR
Keywords:
real-time RT-PCR, virus, SARS-CoV-2, methodAbstract
During the pandemic, key priorities included diagnosis, prevention, and elimination of the disease. Numerous test systems with varying levels of sensitivity were developed for diagnostic purposes. Among these systems, molecular-genetic assays hold a particularly important place, as they enable accurate identification of infectious agents. Within this approach, the most widely used method for diagnostic purposes in scientific research and clinical settings is real-time RT-PCR. In this work, we provide a brief overview of the real-time RT-PCR method. The fundamental principles of the method, its variants, as well as its advantages and limitations, are outlined.
References
COVID-19 Cases, World. https://data.who.int/dashboards/covid19/cases
Carter LJ, Garner LV, Smoot JW, Li Y, Zhou Q, Saveson CJ, Sasso JM, Gregg AC, Soares DJ, Beskid TR, Jervey SR, Liu C. Assay Techniques and Test Development for COVID-19 Diagnosis. ACS Cent Sci. 2020 May 27;6(5):591-605. doi: 10.1021/acscentsci.0c00501
Mackay IM, Arden KE, Nitsche A. Real-time PCR in virology. Nucleic Acids Res. 2002 Mar 15;30(6):1292-305. doi: 10.1093/nar/30.6.1292.
Artika IM, Dewi YP, Nainggolan IM, Siregar JE, Antonjaya U. 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
Haddad F, Baldwin KM. Reverse transcription of the ribonucleic acid: the first step in RT-PCR assay. Methods Mol Biol. 2010;630:261-70. doi: 10.1007/978-1-60761-629-0_17
Saiki RK, Gelfand DH, Stoffel S, Scharf SJ, Higuchi R, Horn GT, Mullis KB, Erlich HA. Primer-directed enzymatic amplification of DNA with a thermostable DNA polymerase. Science. 1988 Jan 29;239(4839):487-91. doi: 10.1126/science.2448875
Chung YS, Lee NJ, Woo SH, Kim JM, Kim HM, Jo HJ, Park YE, Han MG. Validation of real-time RT-PCR for detection of SARS-CoV-2 in the early stages of the COVID-19 outbreak in the Republic of Korea. Sci Rep. 2021 Jul 20;11(1):14817. doi: 10.1038/s41598-021-94196-3.
Mo Y, Wan R, Zhang Q. Application of reverse transcription-PCR and real-time PCR in nanotoxicity research. Methods Mol Biol. 2012;926:99-112. doi: 10.1007/978-1-62703-002-1_7.
Wong ML, Medrano JF. Real-time PCR for mRNA quantitation. Biotechniques. 2005 Jul;39(1):75-85. doi: 10.2144/05391RV01
Arnaudova K.Sh., Yasenyavskaya A.L., Rostoshvili G.A., Samotrueva M.A., Bashkina O.A. Modern molecular-genetic technologies in the diagnosis of viral diseases. RMJ. Medical Review. 2021;5(7):497-502. DOI: 10.32364/2587-6821-2021-5-7-497-502.
Trotskaya O.S. Modern molecular-genetic technologies in the diagnosis of viral diseases.https://www.10gkb.by/uslugi/platnye-mlugi/otdelenie-meditsinskoj-reabilitatsii/sovremennye-molekulyarno-geneticheskie-tekhnologii-v-diagnostike-virusnykh-zabolevanij
Afzal A. Molecular diagnostic technologies for COVID-19: Limitations and challenges. J Adv Res. 2020 Nov;26:149-159. doi: 10.1016/j.jare.2020.08.002
Nguyen NNT, McCarthy C, Lantigua D, Camci-Unal G. Development of Diagnostic Tests for Detection of SARS-CoV-2. Diagnostics (Basel). 2020 Nov 5;10(11):905. doi: 10.3390/diagnostics10110905
Li X, Geng M, Peng Y, Meng L, Lu S. Molecular immune pathogenesis and diagnosis of COVID-19. J Pharm Anal. 2020 Apr;10(2):102-108. doi: 10.1016/j.jpha.2020.03.001