A brief history of RT-PCR and our laboratory experience with SARS-CoV-2 analyses using RT-PCR: RT-PCR and SARS-CoV-2
Polymerase Chain Reaction (PCR) is in vitro replication that allows accelerated amplification of certain sequences in small DNA fragments. A sensitive technique, only traces of DNA are needed for PCR to produce enough copies to be analyzed. In molecular diagnostic laboratories, rRT-PCR technique is applied to find target RNAs for the diagnosis of specific pathogens. Although the rRT-PCR method, which has high specificity and moderate sensitivity, is accepted by WHO as the gold standard test for the confirmation of COVID-19, there are many negative comments about this method that should be considered. While diagnosing SARS-CoV-2, it is possible to say that real-time PCR (RT-PCR) analysis is still valid but not sufficient to quickly distinguish similar infections. For this reason, there is a need for new analysis methods and new RT-PCR studies to be performed with newly developed unique rapid tests.
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- Mullis K, Faloona F, Scharf S, et al. Spesific enzymatic amplification of DNA in vitro: polymerase chain reaction. Cold Spring Harbor Symposia on Quantitative Biology. 1986;51, 263-273.
- Garcia LT, Cristancho LM, Vera EP, et al. A new multiplex-PCR for urinary tract pathogen detection using primer design based on an evolutionary computation method. J Microbiol Biotechnol. 2015;25(10):1714–27.
- Sreejith KR, Ooi CH, Jin J, et al. Digital polymerase chain reaction tech - nology-recent advances and future per - spectives. Lab Chip. 2018;18(24):3717–32.
- Dixon M, Sha S, Stefil M, et al. Is it Time to Say Goodbye to Culture and Sen - sitivity? The Case for Culture-independent Urology. Urology. 2019;136:112–8.
- Garibyan L, Avashia N. Research Techniques Made Simple: Polymerase Chain Reac - tion(PCR). J Invest Dermatol. 2013;133(3).
- Cilloni D, Petiti J, Rosso V, et al. Digital PCR in myeloid malignancies: Ready to replace quantitative PCR? Int J Mol Sci. 2019;20(9).
- Kurkela S, Brown DWG. Molecular diag - nostic techniques. Medicine (Baltimore). 2009;37(10):535–40.
- Quan PL, Sauzade M, Brouzes E. DPCR: A technology review. Sensors (Switzerland). 2018;18(4):1271.
- Mayer G, Muller J, Lunse CE. RNA diagnostics: real-time RT-PCR strategies and promising novel target RNAs. Wiley Interdiscip. Rev. RNA. 2011;2(1):32–41.
- Fu L, Wang B, Yuan T, et al. Clinical characteristics of coronavirus disease 2019 (COVID-19) in China: A systematic review and meta-analysis. J. Infect. 2020;80(6):656–665.
- Alayunt NO, Ozudogru O, Yerlikaya E. First remarkable findings in comparison of patients in Siirt / Turkey in novel coronavirus (Covid-19) pandemic. Medicine Science 2020;9(4):1008-13
- Corman VM, Landt O, Kaiser M, et al. Detection of 2019 novel coronavirus (2019-nCoV) by real-time RT-PCR. Euro. Surveill. 2020;25(3).
- Lippi G, Simundic AM., Plebani M. Potential preanalytical and analytical vulnerabilities in the laboratory diagnosis of coronavirus disease 2019 (COVID-19) Clin. Chem. Lab. Med. 2020;58(7):1070–1076.
- Espy MJ, Uhl JR, Sloan LM, et al. Real-time PCR in clinical microbiology: applications for routine laboratory testing. Clin. Microbiol. Rev. 2006;19(1):165–256.
- Lippi G, Meyer A, Cadamuro J, et al. European Federation of Clinical, P. Laboratory Medicine Working Group for Preanalytical, PREDICT: a checklist for preventing preanalytical diagnostic errors in clinical trials, Clin. Chem. Lab Med. 58(4) (2020) 518–526.
- Tang YW, Schmitz JE, Persing DH, et al. Laboratory Diagnosis of COVID-19: Current Issues and Challenges. J. Clin. Microbiol. 2020;58(6).
- Wölfel R, Corman VM, Guggemos W, et al. Virological assessment of hospitalized patients with COVID-2019. Nature 2020;581(7809):465–469.
- Centers for Disease Control and Prevention. Interim Guidelines for Collecting, Handling, and Testing Clinical Specimens from Persons for Coronavirus Disease 2019 (COVID-19). Updated July 8, 2020. Accessed April 16, 2020.
- Huang P, Liu T, Huang L, et al. Use of chest CT in combination with negative RT-PCR assay for the 2019 novel coronavirus but high clinical suspicion. Radiology. 2020 Apr;295(1):22–23.
- Xie X, Zhong Z, Zhao W, et al. Chest CT for typical 2019-nCoV pneumonia: relationship to negative RT-PCR testing. Radiology. 2020 doi: 10.1148/radiol.2020200343.
- Lee E, Ng M, Khong PL. COVID-19 pneumonia: what has CT taught us? Lancet Infect Dis. 2020;20(4):384–385.
- American College of Radiology ACR Recommendations for the use of Chest Radiography and Computed Tomography (CT) for Suspected COVID-19 Infection. https://www.acr.org/Advocacy-and-Economics/ACR-Position-Statements/Recommendations-for-Chest-Radiography-and-CT-for-Suspected-COVID19-Infection
- The Royal College of Radiologists The role of CT in patients suspected with COVID-19 infection. https://www.rcr.ac.uk/college/coronavirus-covid-19-what-rcr-doing/clinical-information/role-ct-chest/role-ct-patients
- Kanne J, Little B, Chung J, et al. Essentials for radiologists on COVID-19: an update—radiology scientific expert panel. Radiology. 2020 doi: 10.1148/radiol.2020200527.
- Fang Y, Zhang H, Xie J, et al. Sensitivity of chest CT for COVID-19: comparison to RT-PCR. Radiology. 2020 doi: 10.1148/radiol.2020200432.
- Wu J., Wu X., Zeng W., Zeng W., Guo D., Fang Z. Chest CT findings in patients with coronavirus disease 2019 and its relationship with clinical features. Invest Radiol. 2020;55(5):257–261.
- Alayunt NO, Ozudogru O, Cakan E. Psychological and biochemical Impact of Covid-19 pandemic on health workers in Siirt, Turkey. Medicine Science 2021;10(4):1486-91.
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