Clinical Evaluation of Self-Collected Saliva by Quantitative Reverse Transcription-PCR (RT-qPCR), Direct RT-qPCR, Reverse Transcription–Loop-Mediated Isothermal Amplification, and a Rapid Antigen Test To Diagnose COVID-19

Nagura-Ikeda, Mayu; Imai, K.; Tabata, Sakiko; Miyoshi, Kazuyasu; Murahara, Nami; Mizuno, Tsukasa; Horiuchi, Midori; Kato, Kazúo; Imoto, Yoshitaka; Iwata, Maki; Mimura, Satoshi; Ito, Tsuyoshi; Tamura, Kouichi; Kato, Yasuyuki

doi:10.1128/jcm.01438-20

Cited by 383 publications

(405 citation statements)

References 26 publications

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“…2020 ; Iwasaki et al 2020 ; Jamal et al 2020 ; Kim, Lee, et al 2020 ; McCormick-Baw et al 2020 ; Migueres et al. 2020 ; Nagura-Ikeda et al 2020 ; Pasomsub et al 2020 ; Williams et al 2020 ; Wyllie et al 2020 ). Most of these studies reported the results of analyses conducted with small- and medium-sized patient cohorts (i.e., 200 subjects or fewer), although studies with larger cohorts (i.e., about 1,000 subjects) have been recently published ( Caulley et al 2020 ; Zhu, Guo, et al 2020 ).…”

Section: The Detection Of Sars-cov-2 In Salivamentioning

confidence: 99%

“…These methods allow more rapid diagnosis by direct rRT-PCR without RNA extraction. Similarly, other companies have developed rRT-PCR devices that include fully automated commercial systems that can shorten the bench time per sample by nearly 90%, reducing the possibility of mistakes during specimen handling and allowing analysis of a larger number of patients in a shorter time frame ( Chen et al 2020 ; Lippi, Mattiuzzi, et al 2020 ; Nagura-Ikeda et al 2020 ).…”

Section: Molecular-based Tests For the Detection Of Sars-cov-2 In Salmentioning

confidence: 99%

“…With respect to direct rRT-PCR, 2 groups tested this more rapid procedure on salivary samples and noted a sensitivity that was only slightly lower than the sensitivity shown by the standard protocol with RNA extraction ( Nagura-Ikeda et al 2020 ) or even superimposable ( Fukumoto et al 2020 ). These findings demonstrate that the presence of RNases in saliva does not impair such an alternate protocol, which bypasses the classic RNA isolation and purification to reduce the risk of human error during this phase.…”

Section: Molecular-based Tests For the Detection Of Sars-cov-2 In Salmentioning

confidence: 99%

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Diagnostic Salivary Tests for SARS-CoV-2

et al. 2020

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The diagnosis of Severe Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV-2) infection relies on the detection of viral RNA by real-time reverse transcription polymerase chain reaction (rRT-PCR) performed with respiratory specimens, especially nasopharyngeal swabs. However, this procedure requires specialized medical personnel, centralized laboratory facilities, and time to provide results (from several hours up to 1 d). In addition, there is a non-negligible risk of viral transmission for the operator who performs the procedure. For these reasons, several studies have suggested the use of other body fluids, including saliva, for the detection of SARS-CoV-2. The use of saliva as a diagnostic specimen has numerous advantages: it is easily self-collected by the patient with almost no discomfort, it does not require specialized health care personnel for its management, and it reduces the risks for the operator. In the past few months, several scientific papers, media, and companies have announced the development of new salivary tests to detect SARS-CoV-2 infection. Posterior oropharyngeal saliva should be distinguished from oral saliva, since the former is a part of respiratory secretions, while the latter is produced by the salivary glands, which are outside the respiratory tract. Saliva can be analyzed through standard (rRT-PCR) or rapid molecular biology tests (direct rRT-PCR without extraction), although, in a hospital setting, these procedures may be performed only in addition to nasopharyngeal swabs to minimize the incidence of false-negative results. Conversely, the promising role of saliva in the diagnosis of SARS-CoV-2 infection is highlighted by the emergence of point-of-care technologies and, most important, point-of-need devices. Indeed, these devices can be directly used in workplaces, airports, schools, cinemas, and shopping centers. An example is the recently described Rapid Salivary Test, an antigen test based on the lateral flow assay, which detects the presence of the virus by identifying the spike protein in the saliva within a few minutes.

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Section: The Detection Of Sars-cov-2 In Salivamentioning

confidence: 99%

Section: Molecular-based Tests For the Detection Of Sars-cov-2 In Salmentioning

confidence: 99%

Section: Molecular-based Tests For the Detection Of Sars-cov-2 In Salmentioning

confidence: 99%

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Diagnostic Salivary Tests for SARS-CoV-2

et al. 2020

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“…[6][7][8] Direct detection of SARS-CoV-2 has been proposed in scalable, rapidly deployed formats, but often suffers from low sensitivity that limits effectivity in general population testing. 9,10 Thus, the discovery of additional reagents to enable early and rapid SARS-CoV-2 detection and/or neutralization is critical.…”

Section: Introductionmentioning

confidence: 99%

De NovoDiscovery of High Affinity Peptide Binders for the SARS-CoV-2 Spike Protein

Pomplun

Jbara

Quartararo

et al. 2020

Preprint

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The β-coronavirus SARS-CoV-2 has caused a global pandemic. Affinity reagents targeting the SARS-CoV-2 spike protein, the most exposed surface structure of the virus, are of interest for the development of therapeutics and diagnostics. We used affinity selection-mass spectrometry for the rapid discovery of synthetic high affinity peptide binders for the receptor binding domain (RBD) of the SARS-CoV-2 spike protein. From library screening with 800 million synthetic peptides, we identified three sequences with nanomolar affinities (dissociation constants Kd = 80 to 970 nM) for RBD and selectivity over human serum proteins. Picomolar RBD concentrations in biological matrix could be detected using the biotinylated lead peptide in ELISA format. These peptides might associate with the SARS-CoV-2-spike-RBD at a site unrelated to ACE2 binding, making them potential orthogonal reagents for sandwich immunoassays. We envision our discovery as a robust starting point for the development of SARS-CoV-2 diagnostics or conjugates for virus directed delivery of therapeutics.Abstract Figure

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“…The WHO currently recommends RT-PCR testing using nasopharyngeal (NPS) and oropharyngeal swabs (OPS) as gold standard for SARS-CoV-2 diagnosis and for monitoring viral load [ 4 , 5 ]. OF has been suggested as an alternate clinical sample, easy and safer to collect, minimizing exposure of healthcare workers and could be useful for making a diagnosis and measuring SARS-CoV-2 viral load and viral shedding during the course of the illness and convalescence [ 6 , 7 , 8 , 9 , 10 , 11 , 12 , 13 ]. To et al, demonstrated that SARS-CoV-2 was present in OF specimen of 11 out of 12 patients, with viral load being higher during the first week after symptoms onset and declining thereafter, being detectable until 25 days after symptoms onset (DSO) [ 14 , 15 ].…”

Section: Introductionmentioning

confidence: 99%

Frequency and Duration of SARS-CoV-2 Shedding in Oral Fluid Samples Assessed by a Modified Commercial Rapid Molecular Assay

Bordi

Sberna

Lalle

et al. 2020

Viruses

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Background: RT-PCR on nasopharyngeal (NPS)/oropharyngeal swabs is the gold standard for diagnosis of SARS-CoV-2 infection and viral load monitoring. Oral fluid (OF) is an alternate clinical sample, easy and safer to collect and could be useful for COVID-19 diagnosis, monitoring viral load and shedding. Methods: Optimal assay conditions and analytical sensitivity were established for the commercial Simplexa™ COVID-19 Direct assay adapted to OF matrix. The assay was used to test 337 OF and NPS specimens collected in parallel from 164 hospitalized patients; 50 bronchoalveolar lavage (BAL) specimens from a subgroup of severe COVID-19 cases were also analysed. Results: Using Simplexa™ COVID-19 Direct on OF matrix, 100% analytical detection down to 1 TCID50/mL (corresponding to 4 × 103 copies (cp)/mL) was observed. No crossreaction with other viruses transmitted through the respiratory toute was observed. Parallel testing of 337 OF and NPS samples showed highly concordant results (κ = 0.831; 95 % CI = 0.771–0.891), and high correlation of Ct values (r = 0.921; p < 0.0001). High concordance and elevated correlation was observed also between OF and BAL. Prolonged viral RNA shedding was observed up to 100 days from symptoms onset (DSO), with 32% and 29% positivity observed in OF and NPS samples, respectively, collected between 60 and 100 DSO. Conclusions: Simplexa™ COVID-19 Direct assays on OF have high sensitivity and specificity to detect SARS-CoV-2 RNA and provide an alternative to NPS for diagnosis and monitoring SARS-CoV-2 shedding.

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Clinical Evaluation of Self-Collected Saliva by Quantitative Reverse Transcription-PCR (RT-qPCR), Direct RT-qPCR, Reverse Transcription–Loop-Mediated Isothermal Amplification, and a Rapid Antigen Test To Diagnose COVID-19

Cited by 383 publications

References 26 publications

Diagnostic Salivary Tests for SARS-CoV-2

Diagnostic Salivary Tests for SARS-CoV-2

De NovoDiscovery of High Affinity Peptide Binders for the SARS-CoV-2 Spike Protein

Frequency and Duration of SARS-CoV-2 Shedding in Oral Fluid Samples Assessed by a Modified Commercial Rapid Molecular Assay

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