Molecular Detection of SARS-CoV-2 Infection in FFPE Samples and Histopathologic Findings in Fatal SARS-CoV-2 Cases

Am J Clin Pathol. 2020 Jul 7;154(2):190-200. doi: 10.1093/ajcp/aqaa091.

Abstract

Objectives: To report methods and findings of 2 autopsies with molecular evaluation of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) positive individuals.

Methods: Postmortem examination was completed following Centers for Disease Control and Prevention public guidelines. Numerous formalin-fixed paraffin-embedded (FFPE) tissue types from each case were surveyed for SARS-CoV-2 RNA by quantitative reverse transcription polymerase chain reaction (qRT-PCR). SARS-CoV-2 viral genome was sequenced by next-generation sequencing (NGS) from FFPE lung tissue blocks.

Results: Postmortem examinations revealed diffuse alveolar damage, while no viral-associated hepatic, cardiac, or renal damage was observed. Viral RNA was detected in lungs, bronchi, lymph nodes, and spleen in both cases using qRT-PCR method. RNA sequencing using NGS in case 1 revealed mutations most consistent with Western European Clade A2a with ORF1a L3606F mutation.

Conclusions: SARS-CoV-2 testing and viral sequencing can be performed from FFPE tissue. Detection and sequencing of SARS-CoV-2 in combination with morphological findings from postmortem tissue examination can aid in gaining a better understanding of the virus's pathophysiologic effects on human health.

Keywords: Autopsy; COVID-19; FFPE; Molecular testing; SARS-CoV-2; Coronavirus; Viral sequencing; nCov-19.

Publication types

  • Case Reports

MeSH terms

  • Aged, 80 and over
  • Autopsy
  • Betacoronavirus / genetics
  • Betacoronavirus / isolation & purification*
  • COVID-19
  • COVID-19 Testing
  • Clinical Laboratory Techniques / methods*
  • Coronavirus Infections / diagnosis*
  • Coronavirus Infections / pathology
  • Coronavirus Infections / virology
  • Fatal Outcome
  • High-Throughput Nucleotide Sequencing
  • Humans
  • Lung / pathology*
  • Lung / virology
  • Male
  • Middle Aged
  • Molecular Diagnostic Techniques / methods*
  • Pandemics
  • Pneumonia, Viral / diagnosis*
  • Pneumonia, Viral / pathology
  • Pneumonia, Viral / virology
  • RNA, Viral / analysis
  • RNA, Viral / isolation & purification
  • Reverse Transcriptase Polymerase Chain Reaction
  • SARS-CoV-2
  • Sequence Analysis, RNA / methods
  • Tissue Fixation / methods

Substances

  • RNA, Viral