Heat Inactivation of Different Types of SARS-CoV-2 Samples: What Protocols for Biosafety, Molecular Detection and Serological Diagnostics?

Viruses. 2020 Jul 7;12(7):735. doi: 10.3390/v12070735.

Abstract

Standard precautions to minimize the risk of SARS-CoV-2 transmission implies that infected cell cultures and clinical specimens may undergo some sort of inactivation to reduce or abolish infectivity. We evaluated three heat inactivation protocols (56 °C-30 min, 60 °C-60 min and 92 °C-15 min) on SARS-CoV-2 using (i) infected cell culture supernatant, (ii) virus-spiked human sera (iii) and nasopharyngeal samples according to the recommendations of the European norm NF EN 14476-A2. Regardless of the protocol and the type of samples, a 4 Log10 TCID50 reduction was observed. However, samples containing viral loads > 6 Log10 TCID50 were still infectious after 56 °C-30 min and 60 °C-60 min, although infectivity was < 10 TCID50. The protocols 56 °C-30 min and 60 °C-60 min had little influence on the RNA copies detection, whereas 92 °C-15 min drastically reduced the limit of detection, which suggests that this protocol should be avoided for inactivation ahead of molecular diagnostics. Lastly, 56 °C-30 min treatment of serum specimens had a negligible influence on the results of IgG detection using a commercial ELISA test, whereas a drastic decrease in neutralizing titers was observed.

Keywords: COVID-19; ELISA; SARS-CoV-2; coronavirus; heat inactivation; neutralization; serology; virus neutralization test.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Antibodies, Neutralizing / immunology
  • Betacoronavirus* / immunology
  • COVID-19
  • Containment of Biohazards / methods*
  • Containment of Biohazards / standards
  • Coronavirus Infections / diagnosis
  • Coronavirus Infections / prevention & control
  • Coronavirus Infections / virology*
  • Enzyme-Linked Immunosorbent Assay
  • Hot Temperature
  • Humans
  • Neutralization Tests
  • Pandemics / prevention & control
  • Pneumonia, Viral / diagnosis
  • Pneumonia, Viral / prevention & control
  • Pneumonia, Viral / virology*
  • SARS-CoV-2
  • Serologic Tests / methods*
  • Serologic Tests / standards
  • Virus Inactivation*

Substances

  • Antibodies, Neutralizing