Detection of SARS-CoV-2 in saliva: implications for specimen transport and storage

J Med Microbiol. 2021 Feb;70(2):001285. doi: 10.1099/jmm.0.001285.

Abstract

Saliva has recently been proposed as a suitable specimen for the diagnosis of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2). Use of saliva as a diagnostic specimen may present opportunities for SARS-CoV-2 reverse transcription polymerase chain reaction (RT-PCR) testing in remote and low-resource settings. Determining the stability of SARS-CoV-2 RNA in saliva over time is an important step in determining optimal storage and transport times. We undertook an in vitro study to assess whether SARS-CoV-2 could be detected in contrived saliva samples. The contrived saliva samples comprised 10 ml pooled saliva spiked with gamma-irradiated SARS-CoV-2 to achieve a concentration of 2.58×104 copies ml SARS-CoV-2, which was subsequently divided into 2 ml aliquots comprising: (i) neat saliva; and a 1 : 1 dilution with (ii) normal saline; (iii) viral transport media, and (iv) liquid Amies medium. Contrived samples were made in quadruplicate, with two samples of each stored at either: (i) room temperature or (ii) 4 °C. SARS-CoV-2 was detected in all SARS-CoV-2 spiked samples at time point 0, day 1, 3 and 7 at both storage temperatures using the N gene RT-PCR assay and time point 0, day 1 and day 7 using the Xpert Xpress SARS-CoV-2 (Cepheid, Sunnyvale, USA) RT-PCR assay. The ability to detect SARS-CoV-2 in saliva over a 1 week period is an important finding that presents further opportunities for saliva testing as a diagnostic specimen for the diagnosis of SARS-CoV-2.

Keywords: COVID-19; SARS-CoV-2; coronavirus disease; molecular microbiology; saliva.

MeSH terms

  • COVID-19 / diagnosis*
  • COVID-19 Testing / methods*
  • Humans
  • Molecular Diagnostic Techniques
  • RNA, Viral / analysis*
  • Reverse Transcriptase Polymerase Chain Reaction
  • SARS-CoV-2 / genetics*
  • SARS-CoV-2 / isolation & purification
  • Saliva / virology*
  • Specimen Handling

Substances

  • RNA, Viral