eISSN: 2221-6197 DOI: 10.31301/2221-6197

Problematic aspects of diagnostics of SARS-CoV-2 coronavirus infection using reverse-transcriptional PCR

Year: 2020

Pages: 564-590

Number: Volume 12, issue 4

Summary:

The paper considers the problematic aspects of detecting a new coronavirus infection using RT-PCR, which often lead to false negative diagnostic results, which occur both at the preanalytic stage and during nucleic acid amplification, including the interpretation of the obtained data. The viral load in humans was evaluated and assumptions were made about the expected number of viral particles in the studied oropharyngeal and nasopharyngeal samples. The list of diagnostic test systems approved for use in the Russian Federation for detecting SARS-CoV-2 with their brief characteristics is given. The necessity of simultaneous use of several targets in the coronavirus genome in one test system detected by probes with the same fluorochrome is noted, which on the one hand increases the probability of detection by increasing the signal, and on the other hand eliminates the false negative results that could occur in the case of mutations in the virus genome at the sites of annealing primers and hybridization probes.

Keywords:

coronavirus, SARS-CoV-2, RNA, COVID-19, diagnostics, real-time RT-PCR, digital PCR, false negative result, nasopharyngeal swab

References:

  1. Abe T., Ikeda T., Tokuda Y., Ito J., Suzuki Y., Narahara C, Iriyama H., Sato K. A patient infected with SARS-CoV-2 over 100 days. QJM. 2020. doi: 10.1093/qjmed/hcaa296
  2. Alcoba-Florez J., González-Montelongo R., Íñigo-Campos A., de Artola D.G., Gil-Campesino H., The Microbiology Technical Support Team, Ciuffreda L., Valenzuela-Fernández A, Flores C. Fast SARS-CoV-2 detection by RT-qPCR in preheated nasopharyngeal swab samples. J. Infect. Dis. 2020. V. 97. P. 66-68. doi: 10.1016/j.ijid.2020.05.099
  3. Alteri C., Cento V., Antonello M., Colagrossi L., Merli M., Ughi N., Renica S., Matarazzo E., Di Ruscio F., Tartaglione L., Colombo J., Grimaldi C., Carta S., Nava A., Costabile V., Baiguera C., Campisi D.., Fanti D, Vismara C., Fumagalli R., Scaglione F., Epis O.M., Puoti M., Perno C.F. Detection and quantification of SARS-CoV-2 by droplet digital PCR in real-time PCR negative nasopharyngeal swabs from suspected COVID-19 patients. PLoS One. 2020. V. 15(9):e0236311. doi: 10.1371/journal.pone.0236311
  4. Arora R., Goel R., Kumar S., Chhabra M., Saxena S., Manchanda V., Pumma P. Evaluation of SARS-CoV-2 in Tears of Patients with Moderate to Severe COVID-19. 2020. S0161-6420(20)30847-2. doi: 10.1016/j.ophtha.2020.08.029
  5. Artesi M., Bontems S., Göbbels P, Franckh M., Maes P., Boreux R., Meex C., Melin P., Hayette M.P., Bours V., Durkin K. A Recurrent Mutation at Position 26340 of SARS-CoV-2 Is Associated with Failure of the E Gene Quantitative Reverse Transcription-PCR Utilized in a Commercial Dual-Target Diagnostic Assay. Clin. Microbiol. 2020. V. 58(10):e01598-20. doi: 10.1128/JCM.01598-20
  6. Baldacci S., Giannico O.V., Giorgino A., Buccoliero G.B., Desiante F., Fragnelli G.R., Rizzi R., Loconsole D., Centrone F., Chironna M., Conversano M. 63 days detection of SARS-CoV-2 RNA from a recovered patient in Southern Italy: A Case Report. Ig. 2020. V. 32(5). P. 590-592. doi: 10.7416/ai.2020.2378
  7. Barra G.B., Santa Rita T.H., Mesquita P.G., Jácomo R.H., Nery L.F.A. Analytical Sensitivity and Specificity of Two RT-qPCR Protocols for SARS-CoV-2 Detection Performed in an Automated Workflow. (Basel). 2020. V. 11(10):1183. doi: 10.3390/genes11101183
  8. Baymiev An.Kh., Baymiev Al.Kh., Kuluev B.R., Shvets K.Yu., Yamidanov R.S., Matniyazov R.T., Chemeris D.A., Zubov V.V., Alekseev Ya.I., Mavzyutov A.R., Ivanenkov Ya.A., Chemeris A.V. Modern approaches to differentiation of live and dead bacteria using selective amplification of nucleic acids. Microbiology. 2020. V. 89, No. 1. P. 13–27. DOI: 10.1134/S0026261720010038
  9. Becker M.G., Taylor T., Kiazy, S., Cabiles D.R., Meyers A.F., Sandstrom P.A. Recommendations for sample pooling on the Cepheid GeneXpert® system using the Cepheid Xpert® Xpress SARS-CoV-2 assay. PLoS One. 2020. V.15 (11):e0241959. doi.org/10.1371/journal.pone.0241959
  10. Bruijns B.B., Tiggelaar R.M., Gardeniers H. The Extraction and Recovery Efficiency of Pure DNA for Different Types of Swabs. Forensic Sci. 2018. V. 63 (5). P. 1492-1499. doi.org/10.1111/1556-4029.13837
  11. Butler J.M. Fundamentals of forensic DNA typing. Amsterdam, Academic Press/Elsevier. 2010. 500 P.
  12. Chan R.W.Y., Chan K.C., Chan K.Y.Y., Lui G.C.Y., Tsun J.G.S., Wong R.Y.K., Yu M.W.L., Wang M.H.T., Chan P.K.S., Lam H.S., Li A.M. SARS-CoV-2 detection by nasal strips: a superior tool for surveillance of pediatric population. Infect. 2020. S0163-4453(20)30704-0. doi: 10.1016/j.jinf.2020.11.009
  13. Chemeris A.V., Chemeris D.A., Magdanov E.G., Garafutdinov R.R., Nagaev N.R., Vakhitov V.A. Causes of false-negative PCR and how to avoid some of them. Biomics. 2012. V.4. P.31-47. (In Russian)
  14. Chemeris A.V., Magdanov E.G., Garafutdinov R.R., Vakhitov V.A. How to avoid the appearance of false-positive results in a polymerase chain reaction? A. Ovchinnikov Bulletin of Biotechnology and Physical and Chemical Biology. 2012. V. 8(3). P. 34-45. (In Russian)
  15. Chemeris D.A., Magdanov E.G., Mashkov O.I., Garafutdinov R.R., Chemeris A.V. Hot start or time-release PCR. Biomics. 2011. V.2(1). P.1-8. (In Russian)
  16. Dust K., Hedley A., Nichol, K., Stein , Adam  H., Karlowsky J. A., Bullard J., Van Caeseele P., Alexander D. C. Comparison of commercial assays and laboratory developed tests for detection of SARS-CoV-2. J. Virol. Methods. 2020. V. 285. 113970. doi:10.1016/j.jviromet.2020.113970
  17. Freire-Paspuel B., Garcia-Bereguiain M. A. Analytical sensitivity and clinical performance of a triplex RT-qPCR assay using CDC N1, N2, and RP targets for SARS-CoV-2 diagnosis. J. Infect. Dis. 2020. V. 102. P 14-16. doi: org/10.1016/j.ijid.2020.10.047
  18. Fukumoto, T., Iwasaki, S., Fujisawa, S., Hayasaka, K., Sato, K., Oguri, S., TakiK., NakakuboS., KamadaK., YamashitaY., KonnoS., NishidaM., SugitaJ., TeshimaT. Efficacy of a novel SARS-CoV-2 detection kit without RNA extraction and purification. J. Infect. Dis. 2020. V. 98. P. 16-17. doi.org/10.1016/j.ijid.2020.06.074
  19. Garafutdinov R.R., Mavzyutov A.R., Alekseev Ya.I., Vorobiev A.A., Nikonorov Yu.M., Chubukova O.V., Matniyazov R.T., Baimiev An.Kh., Maksimov I.V., Kuluev B.R., Baimiev Al.Kh., Chemeris A.V. Human betacoronaviruses and their highly sensitive detection using PCR and other amplification methods. Biomics. 2020. V.12 (1). P. 121-179. doi: 31301/2221-6197.bmcs.2020-7 (In Russian)
  20. Garafutdinov R.R., Mavzyutov A.R., Nikonorov Yu.M., Chubukova O.V., Matniyazov R.T., Baymiev An.Kh., Maksimov I.V., Miftakhov I.Yu., Khalikova E.Yu., Kuluev B.R., Baymiev Al.Kh., Chemeris A.V. Betacoronavirus SARS-CoV-2, its genome, variety of genotypes and molecular-biological approaches to combat it. Biomics. 2020. V.12(2). P. 242-271. DOI: 10.31301/2221-6197.bmcs.2020-15 (In Russian)
  21. Garg A., Ghosha U., Patel S. S., Singh D. V., Arya A. K., Vasanth S., Pandey A., Srivastava N.. Evaluation of seven commercial RT‐PCR kits for COVID‐19 testing in pooled clinical specimens. Med. Virol. 2020 doi: 10.1002/jmv.26691
  22. Haddar C., Verhoeven P. O., Bourlet T., Pozzetto B., Pillet, S. Brief comparative evaluation of six open one-step RT-qPCR mastermixes for the detection of SARS-CoV-2 RNA using a Taqman probe. Clin. Virol. 2020. 132. P. 104636. 10.1016/j.jcv.2020.104636.
  23. Iwasaki S., Fujisawa S., Nakakubo S., Kamada K., Yamashita Y., Fukumoto T., Sato K., Oguri S., Taki K., Senjo H., Sugita J., Hayasaka K., Konno S., Nishida M., Teshima T.J Comparison of SARS-CoV-2 detection in nasopharyngeal swab and saliva. Journal of Infection. 2020. V. 81(2):e145-e147. doi: 10.1016/j.jinf.2020.05.071
  24. Jacot D., Greub G., Jaton K., Opota O. Viral load of SARS-CoV-2 across patients and compared to other respiratory viruses. Microbes Infect. 2020. V. 22 (10). P. 617-621.doi.org/10.1016/j.micinf.2020.08.004
  25. Jain A., Rophina M., Mahajan S., Krishnan B.B., Sharma M., Mandal S., Fernandez T., Sultanji S., Jolly B., Mathew .S, Sivasubbu S., Scaria V. Analysis of the potential impact of genomic variants in global SARS-CoV-2 genomes on molecular diagnostic assays. J. Infect. Dis. 2020. V. 102. P. 460-462. doi: 10.1016/j.ijid.2020.10.086
  26. Kandel C., Zheng J., McCready J., Serbanescu M.A., Racher H., Desaulnier M., Powis J.E., Vojdani K., Finlay L., Sheldrake E., Vermeiren C., Katz K, McGeer A, Kozak R, Goneau LW. Detection of SARS-CoV-2 from Saliva as Compared to Nasopharyngeal Swabs in Outpatients. Viruses. 2020. V. 12(11):1314. doi: 10.3390/v12111314
  27. Kandetu T.B., Dziuban E.J., Sikuvi K., Beard R.S., Nghihepa R., van Rooyen G., Shiningavamwe A., Katjitae I. Persistence of positive RT-PCR results for over 70 days in two travelers with COVID-19. Disaster Med Public Health Prep. 1-7. doi: 10.1017/dmp.2020.450
  28. Kim S.M., Hwang Y.J., Kwak Y. Prolonged SARS-CoV-2 detection and reversed RT-PCR results in mild or asymptomatic patients. Dis. (Lond). 2020. doi: 10.1080/23744235.2020.1820076
  29. Kim S.Y., Lee J., Sung H., Lee H., Han M.G., Yoo C.K., Lee S.W., Hong K. H. Pooling Upper Respiratory Specimens for Rapid Mass Screening of COVID-19 by Real-Time RT-PCR. Infect. Dis. 2020a. V. 26 (10). P. 2469-2472. doi: 10.3201/eid2610.201955
  30. Kuchinski K.S., Jassem A.N., Prystajecky N.A. Assessing oligonucleotide designs from early lab developed PCR diagnostic tests for SARS-CoV-2 using the PCR_strainer pipeline. Clin. Virol. 2020. V. 131:104581. doi: 10.1016/j.jcv.2020.104581.
  31. 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. Intern. Med. 2020. V. 173(4). P. 262-267. doi: 10.7326/M20-1495
  32. Kudo E., Israelow B., Vogels C. B., Lu P., Wylli A. L., Tokuyama M., Venkataraman A., Brackney D.E., Ott I.M., Petrone M.E., Earnest R., Lapidus S., Muenker M.C., Moore A.J, Casanovas-Massana A., Omer S.B., Dela Cruz C.S., Farhadian S.F., Ko A.I., Grubaugh N.D., Iwasaki A. Detection of SARS-CoV-2 RNA by multiplex RT-qPCR. PLoS Biology. 2020. V. 18(10). e3000867. doi: 10.1371/journal.pbio.3000867
  33. Lampl B. M., Salzberger B. Changing epidemiology of COVID-19. Hyg. Infect. Control. 2020. V.15.doi: 10.3205/dgkh000362
  34. Leung K., Shum M.H., Leung G.M., Lam T.T., Wu J.T. Early transmissibility assessment of the N501Y mutant strains of SARS-CoV-2 in the United Kingdom, October to November 2020. Euro Surveill. 2021. V.26(1). doi: 10.2807/1560-7917.ES.2020.26.1.2002106
  35. Li X., Zai J., Zhao Q, Nie Q., Li Y., Foley B.T., Chaillon A. Evolutionary history, potential intermediate animal host, and cross-species analyses of SARS-CoV-2. Med. Virol. 2020. V. 92(6). P. 602-611. doi: 10.1002/jmv.25731
  36. Li, Y., Yan L.M., Wan L., Xiang T.X., Le A., Liu J.M., PeirisM., PoonL. L.M., ZhangW. Viral dynamics in mild and severe cases of COVID-19. Lancet Infect. Dis. 2020a. doi:10.1016/S1473-3099(20)30232-2
  37. Liotti F. M., Menchinelli G., Marchetti S., Morandotti G. A., Sanguinetti M., Posteraro B., Cattani, P. Evaluating the newly developed BioFire COVID-19 test for SARS-CoV-2 molecular detection. Microbiol. Infect. 2020. doi:10.1016/j.cmi.2020.07.026
  38. Liu C., Shi Q., Peng M., Lu R., Li H., Cai Y., Chen J., Xu J., Shen B. Evaluation of droplet digital PCR for quantification of SARS-CoV-2 Virus in discharged COVID-19 patients. Aging (Albany NY). V 12(21). P. 20997-21003. doi:10.18632/aging.104020
  39. Liu M., Li Q., Zhou J., Ai W., Zheng X., Zeng J., Liu Y., Xiang X., Guo R., Li X., Wu X., Xu H., Jiang L., Zhang H., Chen J., Tian L., Luo J., Luo C. Value of swab types and collection time on SARS-COV-2 detection using RT-PCR assay. Virol. Methods. 2020. 286:113974. doi: 10.1016/j.jviromet.2020.113974
  40. Liu X., Feng J., Zhang Q., Guo D., Zhang L., Suo T.., Hu W, Guo M., Wang X., Huang Z., Xiong Y., Chen G., Chen Y., Lan K. Analytical comparisons of SARS-COV-2 detection by qRT-PCR and ddPCR with multiple primer/probe sets. Microbes Infect. 2020a. V. 9(1). P. 1175-1179. doi: 10.1080/22221751.2020.1772679
  41. Lübke N., Senff T., Scherger S., Hauka S., Andrée M., Adams O., Timm J., Walker A. Extraction-free SARS-CoV-2 detection by rapid RT-qPCR universal for all primary respiratory materials. Clin. Virol. 2020. V. 130. P.104579. doi: 10.1016/j.jcv.2020.104579
  42. Mancini F., Barbanti F., Scaturro M., Errico G., Iacobino A., Bella A., Riccardo F., Marsili G., Stefanelli P., Pezzotti P., Rezza G., Ciervo A. Laboratory management for SARS-CoV-2 detection: a user-friendly combination of the heat treatment approach and rt-Real-time PCR testing. Microbes Infect. 2020. V. (1). P. 1393-1396. doi: 10.1080/22221751.2020.1775500
  43. Mancini F., Barbanti F., Scaturro M., Fontana S., Di Martino A., Marsili G., Puzelli S., Calzoletti L., Facchini M., Di Mario G., Fabiani C., Bella A., Riccardo F., Pezzotti P., Stefanelli P., Rezza G., Ciervo A. Multiplex rt-Real Time PCR assays for diagnostic testing of SARS-CoV-2 and seasonal influenza viruses. A challenge of the phase 3 pandemic setting. Infect. Dis. 2020. doi: org/10.1093/infdis/jiaa658
  44. Mavziutov A.R., Bondarenko V.M., Latkin A.T. Inhibitore oF the Polymerase Chain Reaction. Journal of Microbiology Epidemiology Immunobiology. 2003. № 3. P. 93-98.
  45. Medeiros da Silva R.C., Nogueira Marinho L.C., de Araújo Silva D.N., Costa de Lima K., Pirih F.Q., Luz de Aquino Martins A.R. Saliva as a possible tool for the SARS-CoV-2 detection: A review. Travel Med. Infect. Dis. 2020. V.38:101920. doi: 10.1016/j.tmaid.2020.101920
  46. Molina L.P., Chow S.K., Nickel A., Love J.E. Prolonged Detection of Severe Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV-2) RNA in an Obstetric Patient With Antibody Seroconversion. Gynecol. 2020. V. 136(4). P. 838-841. doi: 10.1097/AOG.0000000000004086
  47. Momynaliev K.T., Ivanov I.V. The nature of false negative results in the detection of SARS-CoV-2 coronavirus by nucleic acid amplification methods. Vestnik Roszdravnadzora. 2020. N.2. C.11-19. DOI: 10.35576/2070-7940-2020-2-11-19 (In Russian)
  48. Moore N. M., Li H., Schejbal D., Lindsley J., Hayden M. K. Comparison of two commercial molecular tests and a laboratory-developed modification of the CDC 2019-nCoV reverse transcriptase PCR assay for the detection of SARS-CoV-2. Clin. Microbiol. 2020. V. 58(8):e00938-20. doi: 10.1128/JCM.00938-20
  49. Moreno-Contreras J., Espinoza M.A., Sandoval-Jaime C., Cantú-Cuevas M.A., Barón-Olivares H., Ortiz-Orozco O.D., Muñoz-Rangel A.V., Hernández-de la Cruz M., Eroza-Osorio C.M., Arias C.F., López S. Saliva Sampling and Its Direct Lysis, an Excellent Option To Increase the Number of SARS-CoV-2 Diagnostic Tests in Settings with Supply Shortages. Clin. Microbiol. 2020. V. 58(10):e01659-20. doi: 10.1128/JCM.01659-20
  50. Munblit D., Nekliudov N.A., Bugaeva P., Blyuss O., Kislova M., Listovskaya E., Gamirova A., Shikhaleva A., Belyaev V., Timashev P., Yavorovskiy A., Bulanova E., Tsareva N., Avdeev S., Kapustina V.A., Pigolkin Y.I., Dankwa E.A., Kartsonaki C., Pritchard M.G., Fomin V., Svistunov A.A., Butnaru D., Glybochko P. StopCOVID cohort: An observational study of 3,480 patients admitted to the Sechenov University hospital network in Moscow city for suspected COVID-19 infection. Infect. Dis. 2020.doi.org/10.1093/cid/ciaa1535
  51. Munster V.J., Feldmann F., Williamson B.N., van Doremalen N., Pérez-Pérez L., Schulz J., Meade-White K., Okumura A., Callison J., Brumbaugh B., Avanzato V.A., Rosenke R., Hanley P.W., Saturday G., Scott D., Fischer E.R., de Wit E. Respiratory disease in rhesus macaques inoculated with SARS-CoV-2. Nature. 2020 Sep;585(7824):268-272. doi: 10.1038/s41586-020-2324-7
  52. Pan Y., Long L., Zhang D., Yuan T., Cui S., Yang P., Wang Q., Ren S. Potential False-Negative Nucleic Acid Testing Results for Severe Acute Respiratory Syndrome Coronavirus 2 from Thermal Inactivation of Samples with Low Viral Loads. Chem. 2020. V. 202066(6). P. 794-801. doi:10.1093/clinchem/hvaa091
  53. Peñarrubia L., Ruiz M., Porco R., Rao S.N., Juanola-Falgarona M., Manissero D., López-Fontanals M., Pareja J. Multiple assays in a real-time RT-PCR SARS-CoV-2 panel can mitigate the risk of loss of sensitivity by new genomic variants during the COVID-19 outbreak. J. Infect. Dis. 2020. V. 97:225-229. doi:10.1016/j.ijid.2020.06.027
  54. Radbel J., Jagpal S., Roy J., Brooks A., Tischfield J., Sheldon M., Bixby C., Witt D., Gennaro M.L., Horton D.B., Barrett E.S., Carson J.L., Panettieri R.A. Jr., Blaser M.J. Detection of Severe Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV-2) Is Comparable in Clinical Samples Preserved in Saline or Viral Transport Medium. Mol. Diagn. 2020. V. 22(7). P. 871-875. doi:10.1016/j.jmoldx.2020.04.209
  55. Rana D.R, Pokhrel N. Sequence mismatch in PCR probes may mask the COVID-19 detection in Nepal. Cell Probes. 2020. V. 53:101599. doi:10.1016/j.mcp.2020.101599
  56. Ransom E.M., Potter R.F., Wallace M.A., Mitchell K.F., Yarbrough M.L., Burnham C.A., Anderson N.W., Parikh B.A. Comparison of Extraction Methods and Thermocyclers for SARS-CoV-2 Molecular Detection Using Clinical Specimens. Clin. Microbiol. 2020. V. 58(10):e01622-20. doi: 10.1128/JCM.01622-20
  57. Riediker M., Tsai D.H. Estimation of Viral Aerosol Emissions From Simulated Individuals With Asymptomatic to Moderate Coronavirus Disease 2019. JAMA Netw. Open. 2020. V. 3(7):e2013807. doi: 10.1001/jamanetworkopen.2020.13807
  58. Rockx B, Kuiken T., Herfst S., Bestebroer T., Lamers M.M., Oude Munnink B.B., de Meulder D., van Amerongen G., van den Brand J., Okba N.M.A., Schipper D., van Run P., Leijten L., Sikkema R., Verschoor E., Verstrepen B., Bogers W., Langermans J., Drosten C., Fentener van Vlissingen M., Fouchier R., de Swart R., Koopmans M., Haagmans B.L. Comparative pathogenesis of COVID-19, MERS, and SARS in a nonhuman primate model. Science. 2020. V.368 (6494). P. 1012-1015.doi:10.1126/science.abb7314
  59. Ryan D. J., Toomey S., Madden S. F., Casey M., Breathnach O. S., Morris P. G., Grogan L., Branagan P., Costello R.W., De Barra E., Hurley K., Gunaratnam C., McElvaney N.G., OBrien M.E., Sulaiman I., Morgan R.K., Hennessy B.T. Use of exhaled breath condensate (EBC) in the diagnosis of SARS-COV-2 (COVID-19). Thorax. 2020. doi: 10.1136/thoraxjnl-2020-215705
  60. Sawano M., Takeshita K., Ohno H., Oka H. A short perspective on a COVID-19 clinical study:‘diagnosis of COVID-19 by RT-PCR using exhale breath condensate samples’. J. Breath Res. 2020. V.14(4). 042003. doi:10.1088/1752-7163/abb99b
  61. Sahajpal N.S., Mondal A.K., Njau A., Ananth S., Jones K., Ahluwalia P.K., Ahluwalia M., Jilani Y., Chaubey A., Hegde M., Kota V., Rojiani A., Kolhe R. Proposal of RT-PCR-Based Mass Population Screening for Severe Acute Respiratory Syndrome Coronavirus 2 (Coronavirus Disease 2019). Mol. Diagn. 2020. V. 22(10). P. 1294-1299. doi: 10.1016/j.jmoldx.2020.07.001
  62. Sakanashi D, Asai N, Nakamura A, Miyazaki N, Kawamoto Y, Ohno T, Yamada A, Koita I, Suematsu H, Hagihara M, Shiota A, Kurumiya A, Sakata M, Kato S, Muramatsu Y, Koizumi Y, Kishino T, Ohashi W, Yamagishi Y, Mikamo H. Comparative evaluation of nasopharyngeal swab and saliva specimens for the molecular detection of SARS-CoV-2 RNA in Japanese patients with COVID-19. Infect. Chemother. 2021. V. 27(1). P. 126-129. doi: 10.1016/j.jiac.2020.09.027
  63. Sender R, Bar-On YM, Flamholz A, Gleizer S, Bernsthein B, Phillips R, Milo R. The total number and mass of SARS-CoV-2 virions in an infected person. medRxiv. 2020. doi: 10.1101/2020.11.16.20232009
  64. Sethuraman N., Jeremiah S.S., Ryo A.. Interpreting Diagnostic Tests for SARS-CoV-2. JAMA. 2020. V. 323(22). P. 2249-2251. doi: 10.1001/jama.2020.8259
  65. Smith, E., Zhen, W., Manji, R., Schron, D., Duong, S., & Berry, G. J. (2020). Analytical and Clinical Comparison of Three Nucleic Acid Amplification Tests for SARS-CoV-2 Detection // J Clin .Microbiol. 2020. V. 58(9):e01134-20. doi: 1128/JCM.01134-20
  66. Srivatsan S., Han P. D., van Raay, K., Wolf C. R., McCulloch D. J., Kim A. E., Brandstetter E., Martin B., Gehring J., Chen W., Kosuri S., KonnickE. Q., LockwoodC. M., RiederM. J., NickersonD. A., ChuY., ShendureJ., Starita L. M. Preliminary support for a “dry swab, extraction free” protocol for SARS-CoV-2 testing via RT-qPCR. bioRxiv. 2020. doi: 10.1101/2020.04.22.056283
  67. Suo T., Liu X., Feng J., Guo M., Hu W., Guo D., Ullah H., Yang Y., Zhang Q., Wang X., Sajid M., Huang Z., Deng L., Chen T., Liu F., Xu K., Liu Y., Zhang Q., Liu Y., Xiong Y., Chen G., Lan K., Chen Y. ddPCR: a more accurate tool for SARS-CoV-2 detection in low viral load specimens. Microbes Infect. 2020. V. 9(1). P. 1259-1268. doi: 10.1080/22221751.2020.1772678
  68. Trypsteen W., Van Cleemput J., Snippenberg W.V., Gerlo S., Vandekerckhove L. On the whereabouts of SARS-CoV-2 in the human body: A systematic review. PLoS Pathog. 2020. V. 16(10):e1009037. doi: 10.1371/journal.ppat.1009037
  69. Ulloa S., Bravo C., Parra B., Ramirez E., Acevedo A., Fasce R., Fernandez J. A simple method for SARS-CoV-2 detection by rRT-PCR without the use of a commercial RNA extraction kit. Virol. Methods. 2020. V. 285. 113960. doi: 10.1016/j.jviromet.2020.113960
  70. Uwamino Y., Nagata M., Aoki W., Fujimori Y, Nakagawa T., Yokota H., Sakai-Tagawa Y., Iwatsuki-Horimoto K., Shiraki T., Uchida S., Uno S., Kabata H., Ikemura S., Kamata H., Ishii M., Fukunaga K., Kawaoka Y., Hasegawa N., Murata M. Accuracy and stability of saliva as a sample for reverse transcription PCR detection of SARS-CoV-2. Clin. Pathol. 2020. doi: 10.1136/jclinpath-2020-206972
  71. Vanaerschot M., Mann S.A., Webber J.T., Kamm J., Bell S.M., Bell J., Hong S.N., Nguyen M.P., Chan L.Y., Bhatt K.D., Tan M., Detweiler A.M., Espinosa A., Wu W., Batson J., Dynerman D., Wadford D.A., Puschnik A.S., Neff N., Ahyong V., Miller S., Ayscue P., Tato C.M., Paul S., Kistler A.L., DeRisi J.L., Crawford E.D. Identification of a Polymorphism in the N Gene of SARS-CoV-2 That Adversely Impacts Detection by Reverse Transcription-PCR. Clin. Microbiol. 2020. V. 59(1):e02369-20. doi:10.1128/JCM.02369-20
  72. Vasudevan H., Xu P., Servellita V., Miller S., Liu L., Gopez A., Chiu C.Y., Abate A.R. Digital droplet PCR accurately quantifies SARS-CoV-2 viral load from crude lysate without nucleic acid purification. medRxiv [Preprint]. 2020. doi: 10.1101/2020.09.02.20186023
  73. Waked R., Makhoul J., Saliba G., Chehata N., Mortada S., Zoghbi A., Choucair J., Haddad E. Are two consecutive negative RT-PCR results enough to rule out COVID-19? New Microbes New Infect. 2020. 37:100750. doi:10.1016/j.nmni.2020.100750
  74. Wang Y., Song W., Zhao Z., Chen P., Liu J., Li C. The impacts of viral inactivating methods on quantitative RT-PCR for COVID-19. Virus Res. 2020. 285:197988. doi: 10.1016/j.virusres.2020.197988
  75. Williamson BN, Feldmann F, Schwarz B, Meade-White K, Porter DP, Schulz J, van Doremalen N, Leighton I, Kwe Yinda C, Pérez-Pérez L, Okumura A, Lovaglio J, Hanley PW, Saturday G, Bosio CM, Anzick S, Barbian K, Cihlar T, Martens C, Scott DP, Munster VJ, de Wit E. Clinical benefit of remdesivir in rhesus macaques infected with SARS-CoV-2. bioRxiv. 2020 Apr 22:2020.04.15.043166. doi: 10.1101/2020.04.15.043166
  76. Yang S., Stanzione N., Uslan D.Z., Garner O.B., de St Maurice A. Clinical and Epidemiologic Evaluation of Inconclusive COVID-19 PCR Results Using a Quantitative Algorithm. J. Clin. Pathol. 2020. 4:aqaa251. doi: 10.1093/ajcp/aqaa251
  77. Yee R., Truong T., Pannaraj P.S., Eubanks N., Gai E., Jumarang J., Turner L., Peralta A., Lee Y., Dien Bard J. Saliva is a Promising Alternative Specimen for the Detection of SARS-CoV-2 in Children and Adults. Clin. Microbiol. 2020. doi: 10.1128/JCM.02686-20
  78. Zhang Y., Wang C., Han M., Ye J., Gao Y., Liu Z., He T., Li T., Xu M., Zhou .L, Zou G., Lu M., Zhang Z. Discrimination of False Negative Results in RT-PCR Detection of SARS-CoV-2 RNAs in Clinical Specimens by Using an Internal Reference. Virol Sin. 2020. doi: 10.1007/s12250-020-00273-8
  79. Zhen W., Berry G. J. Development of a New Multiplex Real-Time RT-PCR Assay for Severe Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV-2) Detection. Mol. Diagn. 2020. V. 22(12). P. 1367-1372. doi: 10.1016/j.jmoldx.2020.09.004
  80. Ziegler K., Steininger P., Ziegler R., Steinmann J., Korn K., Ensser A. SARS-CoV-2 samples may escape detection because of a single point mutation in the N gene. Euro Surveill. 2020. V. 25(39):2001650. doi: 10.2807/1560-7917.ES.2020.25.39.2001650
Download pdf
up
eISSN: 2221-6197 DOI: 10.31301/2221-6197