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Viruses
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Saliva in the Diagnosis of Viral Diseases
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Saliva in the Diagnosis of Viral Diseases

A special issue of Viruses (ISSN 1999-4915). This special issue belongs to the section "Human Virology and Viral Diseases".

Deadline for manuscript submissions: 31 July 2025 | Viewed by 9814

Special Issue Editors

Dr. Anna Rosa Garbuglia

E-Mail Website
Guest Editor
Lab of Virology, Pad Baglivi, INMI L Spallanzani, Via Portuense, 292, 00149 Rome, Italy
Interests: HPV; HIV; viral hepatitis; virus molecular evolution; host–pathogen interaction; zoonoses
Special Issues, Collections and Topics in MDPI journals
Dr. Licia Bordi

E-Mail Website
Guest Editor
Laboratory of Virology, National Institute for Infectious Diseases "L. Spallanzani", Via Portuense 292, 00149 Rome, Italy
Interests: rapid tests for COVID-19; saliva and COVID-19 diagnosis; emerging viruses; host–pathogen interaction
Special Issues, Collections and Topics in MDPI journals

Special Issue Information

Dear Colleagues,

Saliva has long been used as one of the main biological samples for the detection of CMV congenital infection in newborns, for the detection of HPV in patients with head and neck squamous cell carcinoma (HNSCC), for studies of somatic mutations or as biomarker sources (i.e., to detect miRNAs). During the COVID-19 pandemic, scientific evidence emerged indicating that molecular tests performed on saliva have diagnostic sensitivity and specificity comparable to those observed with nasopharyngeal swabs for SARS-CoV-2 RNA detection. Moreover, the presence of IgA and IgG antibodies at the mucosal level has been demonstrated to influence the progression of viral infection and the severity of clinical manifestation. Looking forward, as saliva uniquely contains both respiratory secretions and immunological components, it potentially has wide applications, ranging from clinical diagnostics to post-vaccine disease burden and immunity surveillance. Due to its easy and painless self-collection, saliva represents a suitable alternative sample in community mass screening programs and for longitudinal sampling of hospitalized individuals aimed at monitoring viral load dynamics and treatment response. Recent proteomic studies revealed that 20–30% of the salivary proteome mirrors the plasma proteome, indicating that saliva could represent a potential alternative approach for systemic disease diagnosis. In this Special Issue, we would like to offer the possibility to describe different purification methods and types of sample collection in order to optimize the use of saliva as specimens for the study of biomarkers, pathogen detection and somatic mutation linked to cancer.

Dr. Anna Rosa Garbuglia
Dr. Licia Bordi
Guest Editors

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Keywords

  • saliva
  • diagnosis
  • pathogen detection
  • immunity
  • biomarkers

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Published Papers (6 papers)

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Research

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9 pages, 937 KiB  
Article
Validation of Saliva as the Clinical Specimen Type for a University-Wide COVID-19 Surveillance Program
by Michael L. Farrell, Anton V. Bryksin, Emily Ryan, Jessica Lin, Naima Djeddar, German Khunteev, Benjamin Holton, Miles Paca, Nicholas Speller, James T. Merrill, Ted M. Ross, Robert J. Hogan, Greg Gibson, Andrés J. García and Michael P. Shannon
Viruses 2024, 16(9), 1494; https://doi.org/10.3390/v16091494 - 21 Sep 2024
Viewed by 1174
Abstract
At the beginning of the COVID-19 pandemic, the Georgia Institute of Technology made the decision to keep the university doors open for on-campus attendance. To manage COVID-19 infection rates, internal resources were applied to develop and implement a mass asymptomatic surveillance program. The [...] Read more.
At the beginning of the COVID-19 pandemic, the Georgia Institute of Technology made the decision to keep the university doors open for on-campus attendance. To manage COVID-19 infection rates, internal resources were applied to develop and implement a mass asymptomatic surveillance program. The objective was to identify infections early for proper follow-on verification testing, contact tracing, and quarantine/isolation as needed. Program success depended on frequent and voluntary sample collection from over 40,000 students, faculty, and staff personnel. At that time, the nasopharyngeal (NP) swab, not saliva, was the main accepted sample type for COVID-19 testing. However, due to collection discomfort and the inability to be self-collected, the NP swab was not feasible for voluntary and frequent self-collection. Therefore, saliva was selected as the clinical sample type and validated. A saliva collection kit and a sample processing and analysis workflow were developed. The results of a clinical sample-type comparison study between co-collected and matched NP swabs and saliva samples showed 96.7% positive agreement and 100% negative agreement. During the Fall 2020 and Spring 2021 semesters, 319,988 samples were collected and tested. The program resulted in maintaining a low overall mean positivity rate of 0.78% and 0.54% for the Fall 2020 and Spring 2021 semesters, respectively. For this high-throughput asymptomatic COVID-19 screening application, saliva was an exceptionally good sample type. Full article
(This article belongs to the Special Issue Saliva in the Diagnosis of Viral Diseases)
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6 pages, 604 KiB  
Communication
Saliva Is a Sensitive and Accessible Sample Both for SARS-CoV-2 Detection and for the Evaluation of Treatment Effectiveness in Follow-Up Studies
by Eleonora Lalle, Valentina Mazzotta, Giuseppe Sberna, Lavinia Fabeni, Anna Rosa Garbuglia, Ilaria Mastrorosa, Alessandra D’Abramo, Emanuele Nicastri, Enrico Girardi, Andrea Antinori, Fabrizio Maggi and Licia Bordi
Viruses 2024, 16(7), 1040; https://doi.org/10.3390/v16071040 - 27 Jun 2024
Viewed by 1043
Abstract
Despite emerging evidence indicating that molecular SARS-CoV-2 tests performed on saliva have diagnostic sensitivity and specificity comparable to those observed with nasopharyngeal swabs (NPSs), most in vivo follow-up studies on the efficacy of drugs against SARS-CoV-2 have been performed on NPSs, not considering [...] Read more.
Despite emerging evidence indicating that molecular SARS-CoV-2 tests performed on saliva have diagnostic sensitivity and specificity comparable to those observed with nasopharyngeal swabs (NPSs), most in vivo follow-up studies on the efficacy of drugs against SARS-CoV-2 have been performed on NPSs, not considering saliva as a possible alternative matrix. For this reason, in this study, we used, in parallel, saliva and NPS samples for the detection of SARS-CoV-2 by real-time RT-PCR in patients receiving Tixagevimab/Cilgavimab, Nirmatrelvir/Ritonavir, or Sotrovimab as a treatment against SARS-CoV-2. Our results showed a good correlation between the NPS and saliva samples for each drug; moreover, comparable changes in the cycle threshold (Ct) levels in saliva and NPSs were observed both 7 days and 30 days after treatment, thus confirming that the saliva represents a good matrix for in vivo follow-up studies verifying the effectiveness of treatments against SARS-CoV-2. Full article
(This article belongs to the Special Issue Saliva in the Diagnosis of Viral Diseases)
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11 pages, 276 KiB  
Article
Investigation of Oral Shedding of Torquetenovirus (TTV) in Moderate-to-Severe COVID-19 Hospitalised Patients
by Rafael Antônio Velôso Caixeta, Alexandre Mendes Batista, Matheus Willian Caetano, Michelle Palmieri, Gabriela Schwab, Rodrigo Melim Zerbinati, Andressa Silva Pereira Victor, Camila de Barros Gallo, Tânia Regina Tozetto-Mendoza, Roger Junges, Karem L. Ortega, André Luiz Ferreira Costa, Dmitry José de Santana Sarmento, Débora Pallos, José Angelo Lauletta Lindoso, Simone Giannecchini and Paulo Henrique Braz-Silva
Viruses 2024, 16(6), 831; https://doi.org/10.3390/v16060831 - 24 May 2024
Cited by 2 | Viewed by 1450
Abstract
Background. Torquetenovirus (TTV) is a small DNA virus constituting the human virome. High levels of TTV-DNA have been shown to be associated with immunosuppression and inflammatory chronic disorders. Aim. To assess the possible association between the salivary viral load of TTV-DNA in patients [...] Read more.
Background. Torquetenovirus (TTV) is a small DNA virus constituting the human virome. High levels of TTV-DNA have been shown to be associated with immunosuppression and inflammatory chronic disorders. Aim. To assess the possible association between the salivary viral load of TTV-DNA in patients hospitalised due to COVID-19 and disease severity. Methods. Saliva samples collected from 176 patients infected with SARS-CoV-2 were used to investigate the presence of SARS-CoV-2 and TTV-DNA by use of real-time RT-PCR. Results. The majority of patients were male with severe COVID-19. Presence of SARS-CoV-2 was observed in the saliva of 64.77% of patients, showing TTV-DNA in 55.68% of them. Patients with impaired clinical conditions (p < 0.001), which evolved to death (p = 0.003), showed a higher prevalence of TTV-DNA. The median viral load in patients with severe condition was 4.99 log10 copies/mL, in which those who were discharged and those evolving to death had values of 3.96 log10 copies/mL and 6.27 log10 copies/mL, respectively. A statistically significant association was found between the distribution of TTV-DNA viral load in saliva samples and severity of COVID-19 (p = 0.004) and disease outcomes (p < 0.001). Conclusions. These results indicate that TTV-DNA in saliva could be a useful biomarker of COVID-19 severity and prognosis. Full article
(This article belongs to the Special Issue Saliva in the Diagnosis of Viral Diseases)

Review

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9 pages, 833 KiB  
Review
Saliva Diagnostics in Spaceflight Virology Studies—A Review
by Douglass M. Diak, Brian E. Crucian, Mayra Nelman-Gonzalez and Satish K. Mehta
Viruses 2024, 16(12), 1909; https://doi.org/10.3390/v16121909 - 12 Dec 2024
Cited by 1 | Viewed by 880
Abstract
Many biological markers of normal and disease states can be detected in saliva. The benefits of saliva collection for research include being non-invasive, ease of frequent sample collection, saving time, and being cost-effective. A small volume (≈1 mL) of saliva is enough for [...] Read more.
Many biological markers of normal and disease states can be detected in saliva. The benefits of saliva collection for research include being non-invasive, ease of frequent sample collection, saving time, and being cost-effective. A small volume (≈1 mL) of saliva is enough for these analyses that can be collected in just a few minutes. For “dry” saliva paper matrices, additional drying times (about 30 min) may be needed, but this can be performed at room temperature without the need for freezers and specialized equipment. Together, these make saliva an ideal choice of body fluid for many clinical studies from diagnosis to monitoring measurable biological substances in hospital settings, remote, and other general locations including disaster areas. For these reasons, we have been using saliva (dry as well as wet) from astronauts participating in short- and long-duration space missions for over two decades to conduct viral, stress, and immunological studies. We have also extended the use of saliva to space analogs including bed rest, Antarctica, and closed-chamber studies. Saliva is a biomarker-rich and easily accessible body fluid that could enable larger and faster public health screenings, earlier disease detection, and improved patient outcomes. This review summarizes our lessons learned from utilizing saliva in spaceflight research and highlights the advantages and disadvantages of saliva in clinical diagnostics. Full article
(This article belongs to the Special Issue Saliva in the Diagnosis of Viral Diseases)
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18 pages, 338 KiB  
Review
The Emergence of Saliva as a Diagnostic and Prognostic Tool for Viral Infections
by Nilson Ferreira de Oliveira Neto, Rafael Antônio Velôso Caixeta, Rodrigo Melim Zerbinati, Amanda Caroline Zarpellon, Matheus Willian Caetano, Debora Pallos, Roger Junges, André Luiz Ferreira Costa, Juan Aitken-Saavedra, Simone Giannecchini and Paulo Henrique Braz-Silva
Viruses 2024, 16(11), 1759; https://doi.org/10.3390/v16111759 - 11 Nov 2024
Cited by 2 | Viewed by 2854
Abstract
Saliva has emerged as a promising diagnostic fluid for viral infections, enabling the direct analysis of viral genetic material and the detection of infection markers such as proteins, metabolites, microRNAs, and immunoglobulins. This comprehensive review aimed to explore the use of saliva as [...] Read more.
Saliva has emerged as a promising diagnostic fluid for viral infections, enabling the direct analysis of viral genetic material and the detection of infection markers such as proteins, metabolites, microRNAs, and immunoglobulins. This comprehensive review aimed to explore the use of saliva as a diagnostic tool for viral infections, emphasizing its advantages and limitations. Saliva stands out due to its simplicity and safety in collection, along with the convenience of self-collection without the need for healthcare supervision, while potentially being comparable to urine and blood in terms of effectiveness. Herein, we highlighted the significant potential of saliva in assessing viral loads and diagnosing viral infections, such as herpesviruses, HPV, PyV, TTV, SARS-CoV-2, and MPXV. The detection of viral shedding in saliva underscores its utility in early diagnosis, the monitoring of infection progression, and evaluating treatment responses. The non-invasive nature of saliva collection makes it an appealing alternative to more invasive methods, promoting better patient compliance and facilitating large-scale screening and surveillance. As such, we further highlight current evidence on the use of saliva as a prognostic tool. Although a significant amount of data is already available, further investigations are warranted to more comprehensively assess the added benefit from the utilization of salivary biomarkers in the clinics. Salivary biomarkers show great promise for the early detection and prevention of viral infection complications, potentially improving disease management and control at the population level. Integrating these non-invasive tools into routine clinical practice could enhance personalized healthcare strategies and patient outcomes. Future studies should focus on establishing standardization protocols, validating the accuracy of salivary diagnostics, and expanding clinical research to enhance the diagnostic and monitoring capabilities of salivary biomarkers. Full article
(This article belongs to the Special Issue Saliva in the Diagnosis of Viral Diseases)

Other

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13 pages, 868 KiB  
Brief Report
Prevalence of EBV, HHV6, HCMV, HAdV, SARS-CoV-2, and Autoantibodies to Type I Interferon in Sputum from Myalgic Encephalomyelitis/Chronic Fatigue Syndrome Patients
by Ulf Hannestad, Annika Allard, Kent Nilsson and Anders Rosén
Viruses 2025, 17(3), 422; https://doi.org/10.3390/v17030422 - 14 Mar 2025
Viewed by 1420
Abstract
An exhausted antiviral immune response is observed in myalgic encephalomyelitis/chronic fatigue syndrome (ME/CFS) and post-SARS-CoV-2 syndrome, also termed long COVID. In this study, potential mechanisms behind this exhaustion were investigated. First, the viral load of Epstein–Barr virus (EBV), human adenovirus (HAdV), human cytomegalovirus [...] Read more.
An exhausted antiviral immune response is observed in myalgic encephalomyelitis/chronic fatigue syndrome (ME/CFS) and post-SARS-CoV-2 syndrome, also termed long COVID. In this study, potential mechanisms behind this exhaustion were investigated. First, the viral load of Epstein–Barr virus (EBV), human adenovirus (HAdV), human cytomegalovirus (HCMV), human herpesvirus 6 (HHV6), and severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) was determined in sputum samples (n = 29) derived from ME/CFS patients (n = 13), healthy controls (n = 10), elderly healthy controls (n = 4), and immunosuppressed controls (n = 2). Secondly, autoantibodies (autoAbs) to type I interferon (IFN-I) in sputum were analyzed to possibly explain impaired viral immunity. We found that ME/CFS patients released EBV at a significantly higher level compared to controls (p = 0.0256). HHV6 was present in ~50% of all participants at the same level. HAdV was detected in two cases with immunosuppression and severe ME/CFS, respectively. HCMV and SARS-CoV-2 were found only in immunosuppressed controls. Notably, anti-IFN-I autoAbs in ME/CFS and controls did not differ, except in a severe ME/CFS case showing an increased level. We conclude that ME/CFS patients, compared to controls, have a significantly higher load of EBV. IFN-I autoAbs cannot explain IFN-I dysfunction, with the possible exception of severe cases, also reported in severe SARS-CoV-2. We forward that additional mechanisms, such as the viral evasion of IFN-I effect via the degradation of IFN-receptors, may be present in ME/CFS, which demands further studies. Full article
(This article belongs to the Special Issue Saliva in the Diagnosis of Viral Diseases)
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