Evaluation of the Advanta Dx SARS-CoV-2 RT-PCR Assay, a High-Throughput Extraction-Free Diagnostic Test for the Detection of SARS-CoV-2 in Saliva: A Diagnostic Accuracy Study

Nasopharyngeal swab specimen (NPS) molecular testing is considered the gold standard for SARS-CoV-2 detection. However, saliva is an attractive, noninvasive specimen alternative. The aim of the study was to evaluate the diagnostic accuracy of Advanta Dx SARS-CoV-2 RT-PCR saliva-based assay against paired NPS tested with either NeumoDxTM SARS-CoV-2 assay or Abbott Real Time SARS-CoV-2 assay as the reference method. We prospectively evaluated the method in two settings: a diagnostic outpatient and a healthcare worker screening convenience sample, collected in November–December 2020. SARS-CoV-2 was detected in 27.7% (61/220) of diagnostic samples and in 5% (10/200) of screening samples. Overall, saliva test in diagnostic samples had a sensitivity of 88.5% (77.8–95.3%) and specificity of 98.1% (94.6–99.6%); in screening samples, the sensitivity was 90% (55.5–99.7%) and specificity 100% (98.1–100%). Our data suggests that the Fluidigm Advanta Dx RT-PCR saliva-based assay may be a reliable diagnostic tool for COVID-19 diagnosis in symptomatic individuals and screening asymptomatic healthcare workers.

A Rapid Saliva Test for Monitoring Immune Protection against SARS-CoV-2 and its Variants

Given the ongoing transmission and emergence of SARS-Cov-2 variants globally, it is critical to have a timely assessment on individuals' immune responses as well as population immunity. Important questions such as the durability of COVID-19 immunity or the efficacy of vaccines require large datasets to generate meaningful insights. However, due to the complexity and relatively high-cost of many immunity assays and the needs for blood-drawing specialists, these assays were mostly limited to small-scale clinical studies. Our work demonstrated the potential of a non-invasive, inexpensive and data-driven solution for large-scale immunity surveillance and for predictive modeling of vaccine efficacy. Combining a proprietary saliva processing method and an ultra-sensitive digital detection technology, we were able to rapidly gather information regarding personalized immune response following infection or vaccination, monitor temporal evolution, and optimize predictive models for variant protection.

Accuracy of SARS-CoV-2 Detection in Saliva for COVID-19 Diagnosis: A Systematic Review and Meta-Analysis

Context: There is an unmet clinical need to develop simple, easy, rapid, and accessible testing for the detection of SARS-CoV-2. Recent reports suggested that saliva may be a host for the virus. The existence of SARS-CoV-2 in saliva can be associated with oral manifestations in infected patients. A systematic review was conducted as well as a meta-analysis to evaluate the diagnostic accuracy of detecting SARS-CoV-2 in saliva and investigate the association between positive saliva test and oral manifestations of COVID-19. Evidence acquisition: A literature search in MEDLINE via PubMed, Scopus, Web of Science, and Cochrane was done in June 2020 and updated in February 2021 using relevant keywords. We screened studies for eligibility. The extracted data were analyzed using Meta-Disc software. Results: Eighteen studies were included. Pooled data from eligible studies showed that the sensitivity of diagnosis of SARS-CoV-2 in saliva was 0.86 (95% CI, 0.83–0.89), and the specificity was 0.98 (95% CI, 0.96–0.98). COVID-19 was associated with oral diseases as amblygeustia, dry mouth, dryness, inflammation of the mouth, and enlargement of lymph nodes in the submandibular regions. Conclusions: Our results showed that the saliva has a high accuracy in the detection of SARS-CoV-2.

Evaluation of SARS-CoV-2 rapid antigen diagnostic tests for saliva samples

Background: The COVID-19 pandemic has highlighted the need for rapid, cost effective and easy-to-use diagnostic tools for SARS-CoV-2 rapid antigen detection (RAD) for use in point of care settings or as self-tests, to limit disease transmission. Using saliva samples would further greatly facilitate sample collection, diagnostic feasibility, and mass screening. Objective: We tested two rapid antigen immunochromatographic tests designed for detection of SARS-CoV-2 in saliva: Rapid Response COVID-19 Antigen Rapid Test Cassette for oral fluids (Rapid Response) and DIAGNOS COVID-19 Antigen Saliva Test (DIAGNOS). Evaluation of detection limit was performed with purified SARS-CoV-2 nucleocapsid protein and titrated live SARS-CoV-2 virus and compared to Abbott Panbio COVID-19 Ag Rapid Test (Panbio) designed for nasopharyngeal samples. Sensitivity and specificity were further evaluated on RT-qPCR positive and negative saliva samples from individuals hospitalized with COVID-19 (n=34); and asymptomatic health care personnel (n=20). Results: The limit of detection of the saliva test from DIAGNOS was comparable with the Panbio test and showed higher sensitivity than Rapid Response for both nucleocapsid protein and diluted live viruses. DIAGNOS and Rapid Response further detected seven (47%) and five (33%), respectively, of the 15 RT-qPCR positive saliva samples in individuals hospitalized with COVID-19. Of the 39 RT-qPCR negative samples, all were negative with both tests (specificity 100%; 95% c.i. 0.91-1.00). Only one of the RT-qPCR positive saliva samples (Ct 21.6) contained infectious virus as determined by cell culture and was also positive using the saliva RADs. Conclusion: The results show that the DIAGNOS test exhibit a similar limit of detection as the Panbio RAD and may be an important and easy-to-use saliva RAD complement to detect infectious individuals.

A smartphone-read ultrasensitive and quantitative saliva test for COVID-19

Point-of-care COVID-19 assays that are more sensitive than the current RT-PCR (reverse transcription polymerase chain reaction) gold standard assay are needed to improve disease control efforts. We describe the development of a portable, ultrasensitive saliva-based COVID-19 assay with a 15-min sample-to-answer time that does not require RNA isolation or laboratory equipment. This assay uses CRISPR-Cas12a activity to enhance viral amplicon signal, which is stimulated by the laser diode of a smartphone-based fluorescence microscope device. This device robustly quantified viral load over a broad linear range (1 to 105 copies/μl) and exhibited a limit of detection (0.38 copies/μl) below that of the RT-PCR reference assay. CRISPR-read SARS-CoV-2 (severe acute respiratory syndrome coronavirus 2) RNA levels were similar in patient saliva and nasal swabs, and viral loads measured by RT-PCR and the smartphone-read CRISPR assay demonstrated good correlation, supporting the potential use of this portable assay for saliva-based point-of-care COVID-19 diagnosis.