Evaluation of Self-collected Saliva Samples Without Viral Transport Media for SARS-CoV-2 Testing via RT-PCR and Comparison of Amplicon Sequences Against Commonly Used Primers in Diagnostic Assays

Mil Med. 2024 Aug 19;189(Suppl 3):184-189. doi: 10.1093/milmed/usae070.

Abstract

Introduction: Mass screening for SARS-CoV-2 using nasopharyngeal swabs (NPS) is costly, uncomfortable for patients, and increases the chance of virus exposure to health care workers. Therefore, this study focused on determining if self-collected unpreserved saliva can be an effective alternative to NPS collection in COVID-19 surveillance.

Materials and methods: In this study, patients being tested for SARS-CoV-2 using NPS were asked to provide a saliva sample to compare their results. NPS samples were evaluated for SARS-CoV-2 using BioFire® FilmArray® Torch® or Cepheid® GeneXpert® systems while saliva samples were evaluated using an in-house developed reverse transcriptase polymerase chain reaction (RT-PCR) which targeted the Envelope (E) and Nucleocapsid (N) genes.

Results: Detection of SARS-CoV-2 using self-collected saliva was found to be only slightly less accurate (<5%) than testing using NPS. In addition, initial saliva RT-PCR identified 27 positive subjects, 18 of which provided amplicons sufficient for confirmatory sequencing. The sequencing data showed a genetic shift in the virus within our population sometime between 22 June and July 8, 2021 from Alpha to Delta variant.

Conclusions: The saliva sample collection method identifies the E gene in SARS COVID-2 samples which provides an alternative specimen source to the NPS. This identifies the S gene and ORF1ab. Saliva collection is more convenient to the patient, yields comparable results to NPS collection, and potentially increases Covid-19 surveillance.

MeSH terms

  • Adult
  • COVID-19 Nucleic Acid Testing* / instrumentation
  • COVID-19 Nucleic Acid Testing* / methods
  • COVID-19 Testing / methods
  • COVID-19* / diagnosis
  • COVID-19* / virology
  • Female
  • Humans
  • Male
  • Nasopharynx / virology
  • Reverse Transcriptase Polymerase Chain Reaction / methods
  • SARS-CoV-2* / genetics
  • SARS-CoV-2* / isolation & purification
  • Saliva* / virology
  • Specimen Handling* / methods