4.6 Article

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

Journal

VIRUSES-BASEL
Volume 12, Issue 7, Pages -

Publisher

MDPI
DOI: 10.3390/v12070735

Keywords

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

Categories

Funding

  1. European Virus Archive Global (EVA-GLOBAL) project from the European Union's Horizon 2020-INFRAIA-2019 project [871029]
  2. Advanced Nanosensing platforms for Point of care global diagnostics and surveillance (CONVAT) H2020 Project [101003544]
  3. PREPMedVet (Preparedness and Response in an Emergency contact to Pathogens of Medical and Veterinary importance) within the Agence Nationale de la Recherche Franco-German call on Civil security/Global security 2019 Edition
  4. Viral Hemorrhagic fever modern approaches for developing bedside rapid diagnostics, IMI2 Program, H2020 [823666]
  5. Inserm through the Reacting (REsearch and ACTion Targeing emerging infectious diseases) initiative

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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 degrees C-30 min, 60 degrees C-60 min and 92 degrees 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 Log(10)TCID50 reduction was observed. However, samples containing viral loads > 6 Log(10)TCID(50)were still infectious after 56 degrees C-30 min and 60 degrees C-60 min, although infectivity was < 10 TCID50. The protocols 56 degrees C-30 min and 60 degrees C-60 min had little influence on the RNA copies detection, whereas 92 degrees 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 degrees 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.

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