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Thermal inactivation of COVID-19 specimens improves RNA quality and quantity.

Maral HemmatiMohsen SoosanabadiTahereh GhorashiHadi GhaffariAzadeh VahediElaheh SabbaghianZahra Rasouli NejadAmir SalatiNavid DanaeiParviz Kokhaei
Published in: Journal of cellular physiology (2020)
The rapid spread of coronavirus disease 2019 (COVID-19), a disease caused by severe acute respiratory syndrome coronavirus 2, poses a huge demand for immediate diagnosis. Real-time reverse transcriptase-polymerase chain reaction (rRT-PCR) of nasopharyngeal (NP) and oropharyngeal (OP) swabs have been used to confirm the clinical diagnosis. To avoid the risk of viral-exposure of laboratory workers, thermal inactivation is currently recommended but has unknown effects on the accuracy of the rRT-PCR results. Thirty-six NP/OP specimens were collected from COVID-19 patients and subjected to thermal inactivation (60°C for 30 min) or the RNA extraction processes to activate the form. Here, our data showed that the concentration of extracted-RNA increases upon thermal inactivation compared to the active form (p = .028).  Significantly higher levels of RNA copy number were obtained in inactivated compared to the active samples for both N and ORF1ab genes (p = .009, p = .032, respectively). Thermal inactivation elevated concentration and copy number of extracted-RNA, possibly through viral-capsid degradation and/or nucleoprotein denaturation.
Keyphrases
  • copy number
  • sars cov
  • coronavirus disease
  • respiratory syndrome coronavirus
  • mitochondrial dna
  • genome wide
  • nucleic acid
  • dna methylation
  • electronic health record
  • machine learning
  • ultrasound guided
  • real time pcr