Validation of a Saliva-Based Test for the Molecular Diagnosis of SARS-CoV-2 Infection

Dis Markers. 2022 Jan 7:2022:6478434. doi: 10.1155/2022/6478434. eCollection 2022.

Abstract

Background: Since the beginning of the pandemic, clinicians and researchers have been searching for alternative tests to improve the screening and diagnosis of the SARS-CoV-2 infection. Currently, the gold standard for virus identification is the nasopharyngeal (NP) swab. Saliva samples, however, offer clear, practical, and logistical advantages but due to a lack of collection, transport, and storage solutions, high-throughput saliva-based laboratory tests are difficult to scale up as a screening or diagnostic tool. With this study, we aimed to validate an intralaboratory molecular detection method for SARS-CoV-2 on saliva samples collected in a new storage saline solution, comparing the results to NP swabs to determine the difference in sensitivity between the two tests.

Methods: In this study, 156 patients (cases) and 1005 asymptomatic subjects (controls) were enrolled and tested simultaneously for the detection of the SARS-CoV-2 viral genome by RT-PCR on both NP swab and saliva samples. Saliva samples were collected in a preservative and inhibiting saline solution (Biofarma Srl). Internal method validation was performed to standardize the entire workflow for saliva samples.

Results: The identification of SARS-CoV-2 conducted on saliva samples showed a clinical sensitivity of 95.1% and specificity of 97.8% compared to NP swabs. The positive predictive value (PPV) was 81% while the negative predictive value (NPV) was 99.5%. Test concordance was 97.6% (Cohen's Kappa = 0.86; 95% CI 0.81-0.91). The LoD of the test was 5 viral copies for both samples.

Conclusions: RT-PCR assays conducted on a stored saliva sample achieved similar performance to those on NP swabs, and this may provide a very effective tool for population screening and diagnosis. Collection of saliva in a stabilizing solution makes the test more convenient and widely available; furthermore, the denaturing properties of the solution reduce the infective risks belonging to sample manipulation.

Publication types

  • Observational Study
  • Validation Study

MeSH terms

  • Adult
  • Aged
  • COVID-19 Nucleic Acid Testing / methods*
  • Case-Control Studies
  • Humans
  • Middle Aged
  • Nasopharynx / virology
  • Predictive Value of Tests
  • RNA, Viral / isolation & purification
  • SARS-CoV-2 / genetics
  • Saliva / virology*
  • Specimen Handling / methods

Substances

  • RNA, Viral