Research Article
Volume 16 Issue 8 - 2020
Sensitivity Evaluation of 2019 Novel Coronavirus (SARS-CoV-2) RT-PCR Detection Kits and Strategy to Reduce False Negative
Yunying Zhou1,2,3, Fengyan Pei1,2, Li Wang4, Huailong Zhao5, Huanjie Li1, Mingyu Ji1,2, Weihua Yang1,2, Qingxi Wang1,2, Qianqian Zhao1,2* and Yunshan Wang1,2,3*
1Medical Research and Laboratory Diagnostic Center, Jinan Central Hospital Affiliated to Shandong University, Jinan, Shandong, China
2Microbiology Department, Jinan Central Hospital Affiliated to Shandong first medical university, Jinan, China
3Research Center of Basic Medicine, Jinan Central Hospital Affiliated to Shandong First Medical University, Jinan, China
4Jinan Infectious Disease Hospital, Shandong University, Jinan, Shandong, China
5Jinan Center for Disease Control and Prevention, China
*Corresponding Author: Qianqian Zhao and Yunshan Wang, Medical Research and Laboratory Diagnostic Center, Jinan Central Hospital Affiliated to Shandong University, Jinan, Shandong, China.
Received: May 29, 2020; Published: July 23, 2020




Abstract

An ongoing outbreak of pneumonia associated with SARS-CoV-2 has now been confirmed globally. In absence of effective vaccines, infection prevention and control through diagnostic testing and quarantine is critical. Early detection and differential diagnosis of respiratory infections increases the chances for successful control of COVID-19 disease. The nucleic acid RT-PCR test is regarded as the current standard for molecular diagnosis with high sensitivity. However, the highest specificity confirmation target ORF1ab gene is considered to be less sensitive than other targets in clinical application. In addition, a large amount of recent evidence indicates that the initial missed diagnosis of asymptomatic patients with SARS-CoV-2 and discharged patients with “re-examination positive” may be due to low viral load, and the ability of rapid mutation of SARS-CoV-2 also increases the rate of false negative results. We aimed to evaluate the sensitivity of different nucleic acid detection kits so as to make recommendations for the selection of validation kit, and amplify the suspicious result to be report-able positive by means of continuous amplification, which is of great significance for the prevention and control of the current epidemic and the discharge criteria of low viral load patients.

Keywords: SARS-CoV-2; False Negative; Sensitivity; Low Viral Load; RT-PCR

References

  1. Munster VJ., et al. “A Novel Coronavirus Emerging in China - Key Questions for Impact Assessment”. The New England Journal of Medicine 8 (2020): 692-694.
  2. Zhu N., et al. “A Novel Coronavirus from Patients with Pneumonia in China, 2019”. The New England Journal of Medicine 8 (2020): 727-733.
  3. Wang D., et al. “Clinical Characteristics of 138 Hospitalized Patients With 2019 Novel Coronavirus-Infected Pneumonia in Wuhan, China”. The Journal of the American Medical Association (2020).
  4. Ashour HM., et al. “Insights into the Recent 2019 Novel Coronavirus (SARS-CoV-2) in Light of Past Human Coronavirus Outbreaks”. Pathogens3 (2020): 186.
  5. Lu R., et al. “Genomic characterisation and epidemiology of 2019 novel coronavirus: implications for virus origins and receptor binding”. Lancet10224 (2020): 565-574.
  6. Corman VM., et al. “Detection of 2019 novel coronavirus (2019-nCoV) by real-time RT-PCR”. Euro Surveill 25.3 (2020).
  7. Pan Y., et al. “Viral load of SARS-CoV-2 in clinical samples”. The Lancet Infectious Diseases 4 (2020): 411-412.
  8. Wang Y., et al. “Combination of RT-qPCR testing and clinical features for diagnosis of COVID-19 facilitates management of SARS-CoV-2 outbreak”. Journal of Medical Virology (2020).
  9. Wang XF., et al. “[Retracted: Clinical and epidemiological characteristics of 34 children with 2019 novel coronavirus infection in Shenzhen]”. Zhonghua Er Ke Za Zhi 58 (2020): E008.
  10. Li Z., et al. “Development and Clinical Application of A Rapid IgM-IgG Combined Antibody Test for SARS-CoV-2 Infection Diagnosis”. Journal of Medical Virology (2020).
  11. Xiang F., et al. “Antibody Detection and Dynamic Characteristics in Patients with COVID-19”. Clinical Infectious Diseases (2020).
  12. Zhao J., et al. “Antibody responses to SARS-CoV-2 in patients of novel coronavirus disease 2019”. Clinical Infectious Diseases (2020): 334.
  13. Wang Y., et al. “Cross-reaction of SARS-CoV antigen with autoantibodies in autoimmune diseases”. Cellular and Molecular Immunology 4 (2004): 304-307.
  14. Jiang G., et al. “Application and optimization of RT-PCR in diagnosis of SARS-CoV-2 infection”. Med Rxiv (2020).
  15. Saiki RK., et al. “Primer-directed enzymatic amplification of DNA with a thermostable DNA polymerase”. Science4839 (1988): 487-491.
  16. Peiris JS., et al. “Clinical progression and viral load in a community outbreak of coronavirus-associated SARS pneumonia: a prospective study”. Lancet9371 (2003): 1767-1772.
  17. Zou L., et al. “SARS-CoV-2 Viral Load in Upper Respiratory Specimens of Infected Patients”. The New England Journal of Medicine (2020).
  18. Peeri NC., et al. “The SARS, MERS and novel coronavirus (COVID-19) epidemics, the newest and biggest global health threats: what lessons have we learned?” International Journal of Epidemiology (2020).
  19. Prompetchara E., et al. “Immune responses in COVID-19 and potential vaccines: Lessons learned from SARS and MERS epidemic”. Asian Pacific Journal of Allergy and Immunology 1 (2020): 1-9.
  20. Broughton JP., et al. “Rapid Detection of 2019 Novel Coronavirus SARS-CoV-2 Using a CRISPR-based DETECTR Lateral Flow Assay”. Med Rxiv (2020).
  21. Lamb LE., et al. “Rapid Detection of Novel Coronavirus (COVID-19) by Reverse Transcription-Loop-Mediated Isothermal Amplification”. Med Rxiv (2020).
  22. Yu L., et al. “Rapid colorimetric detection of COVID-19 coronavirus using a reverse transcriptional loop-mediated isothermal amplification (RT-LAMP) diagnostic plat-form: iLACO”. Med Rxiv (2020).
  23. Gao WJ and LM Li “[Advances on presymptomatic or asymptomatic carrier transmission of COVID-19]”. Zhonghua Liu Xing Bing Xue Za Zhi 41 (2020): 485-488.
  24. Li Y., et al. “Stability issues of RT-PCR testing of SARS-CoV-2 for hospitalized patients clinically diagnosed with COVID-19”. Journal of Medical Virology (2020).
  25. Li D., et al. “False-Negative Results of Real-Time Reverse-Transcriptase Polymerase Chain Reaction for Severe Acute Respiratory Syndrome Coronavirus 2: Role of Deep-Learning-Based CT Diagnosis and Insights from Two Cases”. Korean Journal of Radiology 4 (2020): 505-508.
  26. Dong X., et al. “Eleven faces of coronavirus disease 2019”. Allergy (2020).
  27. Angeletti S., et al. “COVID-2019: The role of the nsp2 and nsp3 in its pathogenesis”. Journal of Medical Virolog 6 (2020): 584-588.
  28. Su YC., et al. “Discovery of a 382-nt deletion during the early evolution of SARS-CoV-2”. Bio Rxiv (2020).
  29. Fang Y., et al. “Sensitivity of Chest CT for COVID-19: Comparison to RT-PCR”. Radiology (2020): 200432.
  30. Ai T., et al. “Correlation of Chest CT and RT-PCR Testing in Coronavirus Disease 2019 (COVID-19) in China: A Report of 1014 Cases”. Radiology (2020): 200642.
  31. Lan L., et al. “Positive RT-PCR Test Results in Patients Recovered From COVID-19”. The Journal of the American Medical Association15 (2020): 1502-1503.
  32. National Genomics Data Center Members and Partners. “Database Resources of the National Genomics Data Center in 2020”. Nucleic Acids Research D1 (2020): D24-D33.
  33. Barra GB., et al. “Analytical sensibility and specificity of two RT-qPCR protocols for SARS-CoV-2 detection performed in an automated workflow”. Med Rxiv (2020).
  34. Zhang W., et al. “Molecular and serological investigation of 2019-nCoV infected patients: implication of multiple shedding routes”. Emerging Microbes and Infections 1 (2020): 386-389.
  35. Tang YW., et al. “The Laboratory Diagnosis of COVID-19 Infection: Current Issues and Challenges”. Journal of Clinical Microbiology (2020).
  36. Patel R., et al. “Report from the American Society for Microbiology COVID-19 International Summit, 23 March 2020: Value of Diagnostic Testing for SARS-CoV-2/COVID-19”. MBio2 (2020).
  37. Wikramaratna P., et al. “Estimating false-negative detection rate of SARS-CoV-2 by RT-PCR”. Med Rxiv (2020).
  38. Niu P., et al. “[Development of Novel Multiplex Real-time RT-PCR Assays for Detection of MERS-CoV Infection]”. Bing Du Xue Bao3 (2016): 349-354.
  39. Lin C., et al. “Comparison of throat swabs and sputum specimens for viral nucleic acid detection in 52 cases of novel coronavirus (SARS-Cov-2) infected pneumonia (COVID-19)”. Med Rxiv7 (2020): 1089-1094.
  40. Pachetti M., et al. “Emerging SARS-CoV-2 mutation hot spots include a novel RNA-dependent-RNA polymerase variant”. Journal of Translational Medicine 1 (2020): 179.
  41. Benvenuto D., et al. “Evolutionary analysis of SARS-CoV-2: how mutation of Non-Structural Protein 6 (NSP6) could affect viral autophagy”. Journal of Infection 1 (2020): e24-e27.
  42. Phan T. “Genetic diversity and evolution of SARS-CoV-2”. Infection, Genetics and Evolution 81 (2020): 104260.
  43. Spellberg B., et al. “Community Prevalence of SARS-CoV-2 Among Patients With Influenza like Illnesses Presenting to a Los Angeles Medical Center in March 2020”. The Journal of the American Medical Association (2020).
  44. Xing Q., et al. “Precautions are Needed for COVID-19 Patients with Coinfection of Common Respiratory Pathogens”. Med Rxiv (2020).
  45. Wang M., et al. “Clinical diagnosis of 8274 samples with 2019-novel coronavirus in Wuhan”. Med Rxiv (2020).
  46. Kong WH., et al. “SARS-CoV-2 detection in patients with influenza-like illness”. Nature Microbiology (2020).
  47. Wang M., et al. “Nanopore target sequencing for accurate and comprehensive detection of SARS-CoV-2 and other respiratory viruses”. Med Rxiv (2020).
  48. Maache M., et al. “False-positive results in a recombinant severe acute respiratory syndrome-associated coronavirus (SARS-CoV) nucleocapsid-based western blot assay were rectified by the use of two subunits (S1 and S2) of spike for detection of antibody to SARS-CoV”. Clinical and Vaccine Immunology 3 (2006): 409-414.
  49. Woo PC., et al. “False-positive results in a recombinant severe acute respiratory syndrome-associated coronavirus (SARS-CoV) nucleocapsid enzyme-linked immunosorbent assay due to HCoV-OC43 and HCoV-229E rectified by Western blotting with recombinant SARS-CoV spike polypeptide”. Journal of Clinical Microbiology 12 (2004): 5885-5888.
Citation: Qianqian Zhao and Yunshan Wang., et al. “Sensitivity Evaluation of 2019 Novel Coronavirus (SARS-CoV-2) RT-PCR Detection Kits and Strategy to Reduce False Negative”. EC Microbiology 16.8 (2020): 53-63.

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