Sensitive detection and quantification of SARS-CoV-2 in saliva

Adult Male 0301 basic medicine 2. Zero hunger 0303 health sciences Reverse Transcriptase Polymerase Chain Reaction SARS-CoV-2 Science Q R COVID-19 Middle Aged Viral Load Real-Time Polymerase Chain Reaction Article 3. Good health 03 medical and health sciences Medicine Humans RNA, Viral Female Reagent Kits, Diagnostic Saliva
DOI: 10.1038/s41598-021-91835-7 Publication Date: 2021-06-14T10:03:10Z
ABSTRACT
AbstractSaliva has significant advantages as a test medium for detection of SARS-CoV-2 infection in patients, such as ease of collection, minimal requirement of supplies and trained personnel, and safety. Comprehensive validation in a large cohort of prospectively collected specimens with unknown SARS-CoV-2 status should be performed to evaluate the potential and limitations of saliva-based testing. We developed a saliva-based testing pipeline for detection of SARS-CoV-2 nucleic acids using real-time reverse transcription PCR (RT-PCR) and droplet digital PCR (ddPCR) readouts, and measured samples from 137 outpatients tested at a curbside testing facility and 29 inpatients hospitalized for COVID-19. These measurements were compared to the nasal swab results for each patient performed by a certified microbiology laboratory. We found that our saliva testing positively detects 100% (RT-PCR) and 93.75% (ddPCR) of curbside patients that were identified as SARS-CoV-2 positive by the Emergency Use Authorization (EUA) certified nasal swab testing assay. Quantification of viral loads by ddPCR revealed an extremely wide range, with 1 million-fold difference between individual patients. Our results demonstrate for both community screening and hospital settings that saliva testing reliability is on par with that of the nasal swabs in detecting infected cases, and has potential for higher sensitivity when combined with ddPCR in detecting low-abundance viral loads that evade traditional testing methods.
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