View Article

Abstract

Reverse transcription polymerase chain reaction (RT-PCR) has emerged as the cornerstone of diagnostic efforts in the detection of corona viruses, particularly the formidable SARS-CoV-2 responsible for the COVID-19 pandemic. This review article provides a comprehensive overview of the diverse applications of RT-PCR in corona virus detection. The sensitivity and specificity of RT-PCR render it indispensable in early-stage diagnosis, enabling prompt isolation and treatment of infected individuals. Moreover, RT-PCR's ability to quantitatively analyze viral load allows for dynamic tracking of infection progression and the evaluation of therapeutic interventions. In addition to its clinical utility, RT-PCR plays a pivotal role in population-wide screening and surveillance programs, offering crucial insights for public health interventions. The adaptability of RT-PCR to detect specific genetic markers associated with viral variants is paramount in monitoring viral evolution and understanding its spread. Furthermore, this review highlights the pivotal role of RT-PCR in the development and evaluation of vaccines and antiviral drugs, providing invaluable data for vaccine efficacy studies and immune response assessments.

Keywords

Corona Virus, RT-PCR technique, Covid-19, Vaccines, Antibodies

Reference

  1. World Health Organization (WHO). WHO Director-General's opening remarks at the media briefing on COVID-19. 2020 Mar 11 [internet]. Geneva: [cited 2020 Oct 30]. Available from:https://www.who.int/director-general/speeches/detail/who-director-general-s-opening- remarks-at-the-media-briefing-on-covid-19---11-march-2020
  2. Zhu N, Zhang D, Wang W, Li X, Yang B, Song J, et al. A Novel Coronavirus from Patients with Pneumonia in China, 2019. N Engl J Med. 2020;382:727-733. doi: https://doi.org/10.1056/NEJMoa2001017
  3. Liu J, Liao X, Qian S, Yuan J, Wang F, Liu Y et al. Community transmission of severe acute respiratory syndrome coronavirus 2, Shenzhen, China, 2020. Emerg Infect Dis. 2020 Jun;26(6):1320-1323. doi: https://doi.org/10.3201/eid2606.20023.
  4. Zhang R, Li Y, Zhang AL, Wang Y and Molina MJ. Identifying airborne transmission as the dominant route for the spread of COVID-19. PNAS. 2020 June 30;117(26)14857-14863.doi: https://doi.org/10.1073/pnas.2009637117
  5. Doremalen NV, Morris DH, Holbrook MG, Gamble A, Williamson BN, Tamin A, et al. Aerosol and surface stability of SARS-CoV-2 as compared with SARS-CoV-1. N Engl J Med. 2020:382:1564-1567. doi: https://doi.org/10.1056/NEJMc2004973
  6. Chin AWH, Chu JTS, Perera MRA, Hui KPY, Yen HL, Chan MCW, et al. Stability of SARS-CoV-2 in different environmental conditions. Lancet. 2020;1:E10. doi: https://doi.org/10.1016/S2666-5247(20)30003-3.
  7. Ong SW, Tan YK, Chia PY, Lee TH, Ng OT, Wong MSY, et al. Air, surface. environmental and personal protective equipment contamination by severe acute respiratory syndrome coronavirus 2 (SARSCOV-2) from a symptomatic patient. JAMA. 2020 Apr 28;323(16):1610-1612. doi: https://doi.org/10.1001/jama.2020.3227
  8. Nicas M and Best D. A study quantifying the hand-to-face contact rate and its potential application to predicting respiratory tract infection. J Occup Environ Hygiene. 2008;5:347-352.doi: https://doi.org/10.1080/15459620802003896
  9. Macias AE, Torre A, Moreno-Espinosa S, Leal PE, Bourlon MT and Ruiz-Palacios GM. Controlling the novel A (H1N1) influenza virus: don't touch your face! Journal of Hospital Infection. 2009 Nov 1:73(3):280-281.doi: https://doi.org/10.1016/j.jhin.2009.06.017
  10. Kwok YLA, Gralton J and McLaws M. Face touching: A frequent habit that has implications for hand hygiene. Am J of Infection Control. 2015 Feb 01:43(2):112-114.doi: https://doi.org/10.1016/j.ajic.2014.10.015
  11. COVID-19: Government issues advisory asking people to wear 'homemade face covers. The New Indian Express. 2020 April 04 [internet]. New Delhi: [cited 2020 Oct 30]. Available from: https://www.newindianexpress.com/nation/2020/apr/04/covid-19-government-issues-advisory-asking-people-to-wear-homemade-face-covers-2125687.amp
  12. Centers for Disease Control and Prevention (CDC). How to Protect Yourself and Others. Updated 2020 Nov 4. [internet]. Atlanta: [cited 2020 Nov 30]. Available from https://www.cdc.gov/coronavirus/2019-ncov/prevent-getting-sick/prevention.html
  13. Chen Y, Qin G, Chen J. Xu J. Feng D, Wu X, et al. Comparison of Face-Touching Behaviors Before and During the Coronavirus Disease 2019 Pandemic. JAMA Netw Open. 2020;3(7):e2016924. doi: https://doi.org/10.1001/jamanetworkopen.2020.16924
  14. Sun CB, Wang YY, Liu G and Liu Z. Role of the Eye in Transmitting Human Coronavirus: What We Know and What We Do Not Know. Front Public Health 2020 Apr 24:8:155. doi: https://doi.org/10.3389/fpubh.2020.00155
  15. Ni W, Yang X, Yang D, Bao J, Li R, Xiao Y, et al. Role of angiotensin-converting enzyme 2 (ACE2) in COVID-19. Crit Care. 2020 Jul 13;24:422. doi: https://doi.org/10.1186/s13054-020-03120-0
  16. Temin HM. 1976. Howard M. Temin-biographical. In Le Prix Nobel en 1975, ed. W Odelberg Stockholm: Nobel Found. http://www.nobelprize.org/nobel prizes/medicine/laureates/1975/ temin-bio.html
  17. Stent G. 2007. Phage and the Origins of Molecular Biology. Cold Spring Harbor, NY: Cold Spring Harbor  Lah. Press
  18. Dulbecco R, Vogt M. 1954. Plaque formation and isolation of pure lines with poliomyelitis viruses.     J. Exp. Med. 99:167-82
  19. Dulbecco R. 1964. Transformation of cells in vitro by DNA-containing viruses. JAMA 190:721-26
  20. Baltimore D, Eggers HJ, Franklin RM, Tamm 1. 1963. Poliovirus-induced RNA polymerase and the effects of virus-specific inhibitors on its production. PNAS 49:843-49
  21. Baltimore D. 1964. In vitro synthesis of viral RNA by the poliovirus RNA polymerase. PNAS 51:450-56
  22. Vandenberg, O: Martiny, D; Rochas. O van Belkum, A; Kozlakidis, Z. Considerations for diagnostic COVID-19 tests. Nat. Rev. Microbiol.2020, 14, 1-13.
  23. Gerbalenya, A E.; Baker, SC.; Baric, R.S., de Groot, R.J. Drosten, C.; Gulyaeva, A.A.; Haagman, BL; Lauber, C.;Lentowich, A.M.; Neuman, B.W., et al. The species Severe acute respiratory syndrome- related coronavirus: Classifying 2019-nCoV and naming it SARS-CoV-2. Nat. Microbiol. 2020, 5.
  24. Yan, Y. Chang, L., Wang, L. Laboratory testing of SARS-CoV. MERS-CoV, and SARS-CoV-2 (2019-CoV): Current status challenges, and countermeasures. Rev. Mod. Viral 2020, 30, e2106.
  25. Padoan, A., Bonfante, F: Pagliari, M.; Bortolami, A.; Negrini, D.; Zuin, S.; Bozzato, D: Cosma, C., Sciacovelli, L.; Plebani, M. Analytical and clinical performances of five immunoassays for the detection of SARS-CoV-2 antibodies in comparison with neutralization activity. ElioMedicine 2020, 62, 103101.
  26. Sheridan, C. Coronavirus and the race to distribute reliable diagnostics. Nat. Esotecional, 2020. 38, 382-384.
  27. Oran D.P., Topol E.J. Prevalence of Asymptomatic SARS-CoV-2 Infection: A Narrative Review. Ann. Intern. Med. 2020;173:362–367. doi: 10.7326/M20-3012. [PMC free article] [PubMed] [CrossRef] [Google Scholar]
  28. Richardson S., Hirsch J.S., Narasimhan M., Crawford J.M., McGinn T., Davidson K.W., The Northwell COVID-19 Research Consortium. Barnaby D.P., Becker L.B., Chelico J.D., et al. Presenting Characteristics, Comorbidities, and Outcomes Among 5700 Patients Hospitalized With COVID-19 in the New York City Area. JAMA. 2020;323:2052–2059. doi: 10.1001/jama.2020.6775. [PMC free article] [PubMed] [CrossRef] [Google Scholar] 
  29. Lippi, G.; Simundic, A.M.; Plebani, M. Potential preanalytical and                  analytical vulnerabilities in the laboratory diagnosis of coronavirus disease 2019 (COVID-19). Clin. Chem. Lab. Med. 2020, 58, 1070–1076.
  30. Interim Guidelines for Collecting, Handling, and Testing Clinical Specimens for COVID-19.
  31. Irving, S.A.; Vandermause, M.F.; Shay, D.K.; Belongia, E.A. Comparison of nasal and nasopharyngeal swabs for influenza detection in adults. Clin. Med. Res. 2012, 10, 215–218.
  32. Kim, J.Y.; Ko, J.H.; Kim, Y.; Kim, Y.J.; Kim, J.M.; Chung, Y.S.; Kim, H.M.; Han, M.G.; Kim, S.Y.; Chin, B.S. Viral Load Kinetics of SARS-CoV-2 Infection in First Two Patients in Korea. J. Korean Med. Sci. 2020, 35, e86.
  33. Sethuraman, N.; Jeremiah, S.S.; Ryo, A. Interpreting Diagnostic Tests for SARS-CoV-2. JAMA 2020, 323, 2249–2251.
  34. Kucirka, L.M.; Lauer, S.A.; Laeyendecker, O.; Boon, D.; Lessler, J. Variation in False-Negative Rate of Reverse Transcriptase Polymerase Chain Reaction-Based SARS-CoV-2 Tests by Time Since Exposure. Ann. Intern. Med. 2020, 173, 262–267.
  35. Poon, L.L.; Chan, K.H.; Wong, O.K.; Yam, W.C.; Yuen, K.Y.; Guan, Y.; Lo, Y.M.; Peiris, J.S. Early diagnosis of SARS coronavirus infection by real time RT-PCR. J. Clin. Virol. 2003, 28, 233–238.
  36. de Sousa, R.; Reusken, C.; Koopmans, M. MERS coronavirus: Data gaps for laboratory preparedness. J. Clin. Virol. 2014, 59, 4–11.
  37. Kobayashi, T.; Jung, S.M.; Linton, N.M.; Kinoshita, R.; Hayashi, K.; Miyama, T.; Anzai, A.; Yang, Y.; Yuan, B.; Akhmetzhanov, A.R.; et al. Communicating the Risk of Death from Novel Coronavirus Disease (COVID-19). J. Clin. Med. 2020, 9, 580.
  38. Loeffelholz, M.J.; Tang, Y.W. Laboratory diagnosis of emerging human coronavirus infections-The state of the art. Emerg Microbes Infect. 2020, 9, 747-756.
  39. MacKay, M.J., Hooker, AC Afshinnekoa, E., Salit, M., Kelly, J.; Feldstein, J.V., Haft, N., Schenkel, D. Nambi, S., Cai, Y., et al. The  COVID-19 XPRIZE and the need for scalable, fast, and widespread testing. Nat. Biotechnol. 2020, 38, 1021-1024.
  40. Amaout, R. Lee, R.A., Lee, G.R., Callahan, C. Yen, C.F. Senith, K.P, Arora, R., Kirby, JE. SARS-CoV2 Testing: The Limit of Detection Matters. bioRxiv 2020.
  41. Nalla, A.K., Caste, A.M.; Huang, M.W.; Perchetti, G.A.; Sampoleo, R., Shrestha, L.; Wei, Y.; Zhu, H.; Jerome, K.R.; Greninger, A.L Comparative Performance of SARS-CoV-2 Detection Assays Using Seven Different Primer-Probe Sets and One Assay Kit. J. Clin. Microbial. 2020, 58, 00557-20.
  42. Lee, S.H. Testing for SARS-CoV-2 in cellular components by routine nested RT-PCR followed by DNA sequencing, Int. J. Geriatr. Rehabil. 2020, 2, 69-96.
  43. Drosten, C., S. Günther, W. Presier, S. Werf, H. Brodt, S. Becker, et al. 2003. Identification of a novel coronavirus in patients with severe acute respiratory syndrome. N. Engl. J. Med. 348:1967-1976. [PubMed] [Google Scholar]
  44. Poon, L., O. Wong, W. Luk, K. Yuen, J. Peiris, and Y. Guan. 2003. Rapid diagnosis of a coronavirus associated with severe acute respiratory syndrome (SARS). Clin. Chem. 49:953-955. [PMC free article] [PubMed] [Google Scholar]
  45. Dutta G., Kim S., Park S., Yang H. Washing-free heterogeneous immunosensor using proximity-dependent electron mediation between an enzyme label and an electrode. Anal. Chem. 2014;86:4589. doi: 10.1021/ac5006487. [PubMed] [CrossRef] [Google Scholar]
  46. Dutta G., Park S., Singh A., Seo J., Kim S., Yang H. Low-interference washing-free electrochemical immunosensor using glycerol-3-phosphate dehydrogenase as an enzyme label. Anal. Chem. 2015;87:3574. doi: 10.1021/ac504485a. [PubMed] [CrossRef] [Google Scholar]
  47. Li Z., Yi Y., Luo X., Xiong N., Liu Y., Li S., Chen L., Ye F. Development and clinical application of a rapid IgM-IgG combined antibody test for SARS-CoV-2 infection diagnosis. J. Med. Virol. 2020;92:1518–1524. doi: 10.1002/jmv.25727. [PMC free article] [PubMed] [CrossRef] [Google Scholar]

Photo
Sahil Agrawal
Corresponding author

Department of Microbiology, Ashokrao Mane Institute of Pharmacy Ambap-416112, India.

Photo
B. Potdar
Co-author

Department of Microbiology, Ashokrao Mane Institute of Pharmacy, Ambap-416112, India.

Photo
J. Shaikh
Co-author

Department of Microbiology, Ashokrao Mane Institute of Pharmacy, Ambap-416112, India.

Photo
K. Pimpale
Co-author

Department of Microbiology, Ashokrao Mane Institute of Pharmacy, Ambap-416112, India.

Photo
L. Kochage
Co-author

Department of Microbiology, Ashokrao Mane Institute of Pharmacy, Ambap-416112, India.

Photo
N. Chougule
Co-author

Department of Microbiology, Ashokrao Mane Institute of Pharmacy, Ambap-416112, India.

S. Agrawal*, B. Potdar, J. Shaikh, K. Pimpale, L. Kochage, N. Chougule, Detection of Corona Virus: A Review on Applications of RT-PCR in Detection of Corona Virus (Covid-19), Int. J. in Pharm. Sci., 2023, Vol 1, Issue 11, 260-271. https://doi.org/10.5281/zenodo.10124285

More related articles
Biological Activities Of Ginseng And Its Applicati...
Tushar Dewangan , Suraj Sahu, Suraj Patel, Miss Divyani Soni, Dr....
A Comprehensive Review on Analytical Methods for T...
Krutanjali Nikumbh, Kiran Dhamak, ...
Pharmacognostical, Phytochemical Characterisation ...
S. Jasbin Nisha, Abish Mobin M., Deepthi Dileep, Thomas R., Danie...
Exploring Formulation Strategies And Assessing Efficacy: A Study On Hand Wash De...
Pallavi N. Somthane, Vedantika R. Salunkhe, Vedant Mulange, Vighnesha Gawari, ...
Use of Potato and Curry Leaves for Anti-Ageing Cream ...
Aayushi Yelgurwar , Chhavi Rahangdale , Divyani Dahake, Harshada Borekar, Mitali Bodhankar , ...
A Review Of The Impact Of Different Herbs As An Anti Asthmatics...
Shyam Rangari, Divyanshu Gahane, Aishwarya Deshmukh, Milind Umekar, ...
Related Articles
Formulation and Evaluation of Herbal Liquid Toner by Using Cymbopogon citratus a...
Deepa Chandrawanshi, Rutu Chaudhari , Tanu Hatel , Sayli Turkar , Swati Meshram , ...
An Overview on Controlled Porosity Osmotic Tablet...
Saba Rajjak Shaikh, Pradnya K. Gnagurde, Shradha G. Kandalkar, Kajal A. Choursiya, Sheetal B. Gondka...
Industrial Attachment Report On Royal Pharmaceuticals Company...
Sarita Rauniyar, Pradeep Kumar, Anil Gupta, Krishna Thakur, Ranjit Yadav, ...
Review On Abutilion Indicum (Mudra)...
Lahoo Balu Palave , Snehal Kothavale , Mahendra Shendkar, Leena parkhi, Lalit Ambhore , Manish Bhosl...
Biological Activities Of Ginseng And Its Application To Human Health...
Tushar Dewangan , Suraj Sahu, Suraj Patel, Miss Divyani Soni, Dr. Shruti Rathore, ...
More related articles
Biological Activities Of Ginseng And Its Application To Human Health...
Tushar Dewangan , Suraj Sahu, Suraj Patel, Miss Divyani Soni, Dr. Shruti Rathore, ...
Pharmacognostical, Phytochemical Characterisation and Formulative Study of Trich...
S. Jasbin Nisha, Abish Mobin M., Deepthi Dileep, Thomas R., Daniel Xavier Prasad, Surabhi G. S., Pra...
Biological Activities Of Ginseng And Its Application To Human Health...
Tushar Dewangan , Suraj Sahu, Suraj Patel, Miss Divyani Soni, Dr. Shruti Rathore, ...
Pharmacognostical, Phytochemical Characterisation and Formulative Study of Trich...
S. Jasbin Nisha, Abish Mobin M., Deepthi Dileep, Thomas R., Daniel Xavier Prasad, Surabhi G. S., Pra...