Persistence of SARS-CoV-2 in saliva: Implications for late-stage diagnosis and infectious duration

Saliva has been a COVID-19 diagnostic specimen of interest due to its simple collection, scalability, and yield. Yet COVID-19 testing and estimates of the infectious period remain largely based on nasopharyngeal and nasal swabs. We sought to evaluate whether saliva testing captured prolonged presence of SARS-CoV-2 and potential infectiousness later in the disease course. We conducted an observational study of symptomatic COVID-19 patients at University Hospital in Newark, NJ. Paired saliva and nasal specimens from 96 patients were analyzed, including longitudinal analysis of paired observations from 28 of these patients who had multiple time-points. Saliva detected significantly more cases of COVID-19 beyond 5 days (86.1% [99/115] saliva vs 48.7% [56/115] nasal, p-value < 0.001), 9 days (79.4% [50/63] saliva vs 36.5% [23/63] nasal, p-value < 0.001) and 14 days (71.4% [20/28] saliva vs 32.1% [9/28] nasal, p-value = 0.010) of symptoms. Additionally, saliva yielded lower cycle thresholds across all time periods, indicative of higher viral loads in saliva. In the longitudinal analysis, a log-rank analysis indicated that the survival curve for saliva was significantly different from the curve for nasal swabs (p<0.001) with a median survival time for saliva of 18 days compared to 13 days for nasal swabs. We additionally performed saliva viral cultures among a similar COVID-19 patient cohort and noted patients with positive saliva viral cultures between 7 to 28 days of symptoms. Findings from this study suggest that SARS-CoV-2 RNA persists longer and in higher abundance in saliva compared to nasal swabs, with potential of prolonged propagating virus. Testing saliva may thus increase yield for detecting potentially infectious virus even beyond the first five days of symptomatic COVID-19.

[1]  M. Siedner,et al.  Duration of Shedding of Culturable Virus in SARS-CoV-2 Omicron (BA.1) Infection , 2022, The New England journal of medicine.

[2]  K. McPhaul,et al.  Comparison of Saliva and Midturbinate Swabs for Detection of SARS-CoV-2 , 2022, Microbiology spectrum.

[3]  D. Alland,et al.  RT-PCR negative COVID-19 , 2022, BMC Infectious Diseases.

[4]  R. Peeling,et al.  Diagnostics for COVID-19: moving from pandemic response to control , 2021, The Lancet.

[5]  D. Alland,et al.  Sample collection and transport strategies to enhance yield, accessibility, and biosafety of COVID-19 RT-PCR testing , 2021, Journal of medical microbiology.

[6]  J. Bender,et al.  Change in Saliva RT-PCR Sensitivity Over the Course of SARS-CoV-2 Infection. , 2021, JAMA.

[7]  O. Pybus,et al.  Viral infection and transmission in a large, well-traced outbreak caused by the SARS-CoV-2 Delta variant , 2021, Nature communications.

[8]  Å. Nilsdotter-Augustinsson,et al.  Evaluation of SARS-CoV-2 rapid antigen diagnostic tests for saliva samples , 2021, Heliyon.

[9]  H. Kunishima,et al.  Back to normal; serological testing for COVID‐19 diagnosis unveils missed infections , 2021, Journal of medical virology.

[10]  Amjad Husain A novel approach to minimize the false negative COVID-19 diagnosis by inclusion of specific cell markers and multiple sample collection , 2021, MethodsX.

[11]  M. Tan,et al.  Saliva is more sensitive than nasopharyngeal or nasal swabs for diagnosis of asymptomatic and mild COVID-19 infection , 2021, Scientific Reports.

[12]  O. Kharbanda,et al.  Exploring salivary diagnostics in COVID-19: a scoping review and research suggestions , 2021, BDJ open.

[13]  M. M. van der Eerden,et al.  Duration and key determinants of infectious virus shedding in hospitalized patients with coronavirus disease-2019 (COVID-19) , 2021, Nature communications.

[14]  J. Klausner,et al.  Self-Collected Oral Fluid and Nasal Swab Specimens Demonstrate Comparable Sensitivity to Clinician-Collected Nasopharyngeal Swab Specimens for the Detection of SARS-CoV-2 , 2020, Clinical infectious diseases : an official publication of the Infectious Diseases Society of America.

[15]  Elizabeth B White,et al.  Saliva or Nasopharyngeal Swab Specimens for Detection of SARS-CoV-2 , 2020, The New England journal of medicine.

[16]  S. Crovella,et al.  Is FURIN gene expression in salivary glands related to SARS-CoV-2 infectivity through saliva? , 2020, Journal of Clinical Pathology.

[17]  Chengfeng Lei,et al.  On the Calculation of TCID50 for Quantitation of Virus Infectivity , 2020, Virologica Sinica.

[18]  Zachary Schiffman,et al.  Predicting infectious SARS-CoV-2 from diagnostic samples , 2020, Clinical infectious diseases : an official publication of the Infectious Diseases Society of America.

[19]  N. Low,et al.  False-negative results of initial RT-PCR assays for COVID-19: A systematic review , 2020, medRxiv.

[20]  C. Hung,et al.  Prolonged virus shedding even after seroconversion in a patient with COVID-19 , 2020, Journal of Infection.

[21]  P. Vollmar,et al.  Virological assessment of hospitalized patients with COVID-2019 , 2020, Nature.

[22]  W. Siqueira,et al.  Coronavirus COVID-19 impacts to dentistry and potential salivary diagnosis , 2020, Clinical Oral Investigations.

[23]  Bernard Rosner,et al.  Wilcoxon Rank-Based Tests for Clustered Data with R Package clusrank , 2017, J. Stat. Softw..

[24]  N A Obuchowski,et al.  On the comparison of correlated proportions for clustered data. , 1998, Statistics in medicine.