Feasibility of Rapid Diagnostic Technology for SARS-CoV-2 Virus Using a Trace Amount of Saliva

Tokuyama-Toda, Reiko; Muraoka, Masaaki; Terada-Ito, Chika; Ide, Shinji; Horiuchi, Toshikatsu; Amemiya, Takayuki; Fukuoka, Airi; Hamada, Y.; Sejima, Shunsuke; Satomura, Kimio

doi:10.3390/diagnostics11112024

Cited by 3 publications

(14 citation statements)

References 49 publications

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“…One-step RT-PCR was performed using a PCR1100 device. The procedure of the PCR1100 device used was based on previously reported procedures [ 25 ]. In a 20-microliter reaction solution containing 1× Reaction Buffer, 0.25 or 0.50 µL of RT Enzyme Mix, 1.0 µL DNA Polymerase (THUNDERBIRD™ Probe One-step qRT-PCR Kit, TOYOBO Co. Ltd., Osaka, Japan), and a final concentration of primer/probe for each target, 3 µL of a sample (PBS containing human coronavirus 229E washed out from treated non-woven fabric filters) was amplified ( Table 1 ).…”

Section: Methodsmentioning

confidence: 99%

Surface Functionalization of Non-Woven Fabrics Using a Novel Silica-Resin Coating Technology: Antiviral Treatment of Non-Woven Fabric Filters in Surgical Masks

Tsutsumi-Arai

Iwamiya²,

Hoshino

et al. 2022

IJERPH

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Masks are effective for preventing the spread of COVID-19 and other respiratory infections. If antimicrobial properties can be applied to the non-woven fabric filters in masks, they can become a more effective countermeasure against human-to-human and environmental infections. We investigated the possibilities of carrying antimicrobial agents on the fiber surfaces of non-woven fabric filters by applying silica-resin coating technology, which can form silica-resin layers on such fabrics at normal temperature and pressure. Scanning electron microscopy and electron probe microanalysis showed that a silica-resin layer was formed on the fiber surface of non-woven fabric filters. Bioassays for coronavirus and quantitative reverse transcription-polymerase chain reactions (RT-PCR) revealed that all antimicrobial agents tested loaded successfully onto non-woven fabric filters without losing their inactivation effects against the human coronavirus (inhibition efficacy: >99.999%). These results indicate that this technology could be used to load a functional substance onto a non-woven fabric filter by vitrifying its surface. Silica-resin coating technology also has the potential of becoming an important breakthrough not only in the prevention of infection but also in various fields, such as prevention of building aging, protection of various cultural properties, the realization of a plastic-free society, and prevention of environmental pollution.

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Section: Methodsmentioning

confidence: 99%

Surface Functionalization of Non-Woven Fabrics Using a Novel Silica-Resin Coating Technology: Antiviral Treatment of Non-Woven Fabric Filters in Surgical Masks

Tsutsumi-Arai

Iwamiya²,

Hoshino

et al. 2022

IJERPH

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“…Samples were prepared as described in a previous study [ 20 ]. In brief, SARS-CoV-2 positive control RNA samples developed according to the National Institute of Infectious Diseases (NIID) protocol were provided by the NIID (for N primer probe Ver.…”

Section: Methodsmentioning

confidence: 99%

“…Primers and probes were prepared as described in a previous study [ 20 ]. Based on the CDC 2020 protocol for SARS-CoV-2 [ 21 ], oligonucleotide primers and probes for quantitative reverse transcriptase PCR (qRT-PCR) were synthesized to target the N1 or N2 region by Nihon Gene Research Laboratories Inc. (Sendai, Japan) [ 20 ]. Based on the CDC protocol, oligonucleotide primers and probes were also synthesized to detect human RNase P.…”

Section: Methodsmentioning

confidence: 99%

“…For example, the turnaround time is long, a laboratory engineer with specialized knowledge/technique is needed, a relatively large benchtop device is required, and sample collection is complicated and involves a risk of infection to examiners [ 19 ]. Therefore, we developed and reported a new rapid diagnostic technology using a trace amount of saliva, using the PCR1100, which is a compact, lightweight, and portable qPCR device [ 20 ]. This method enabled us to diagnose patients in a short time of about 18 min on average and has the advantage of a much lower risk of infection due to the use og a mouthwash without any pretreatment of the sample.…”

Section: Introductionmentioning

confidence: 99%

“…In this diagnostic procedure, N1 and N2 regions of the SARS-CoV-2 were targeted for quantitative gene amplification using specific primers and probes as recommended by the Centers for Disease Control and Prevention (CDC). Despite successfully amplifying the CDC N1 region during the diagnostic process in infected patients, the detection sensitivity of the N2 region was dramatically low, and CDC N1 was not detected in all cases [ 20 ]. Therefore, this study aimed to improve the detection sensitivity of CDC N2 and established qPCR conditions with higher sensitivity by optimizing annealing conditions and modifying polymerase chain reaction (PCR) reagents.…”

Section: Introductionmentioning

confidence: 99%

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Improving the Detection Sensitivity of a New Rapid Diagnostic Technology for Severe Acute Respiratory Syndrome Coronavirus 2 Using a Trace Amount of Saliva

Tokuyama-Toda

Terada-Ito

Muraoka³

et al. 2022

Diagnostics

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The early diagnosis and isolation of infected individuals with coronavirus disease 2019 (COVID-19) remain important. Although quantitative polymerase chain reaction (qPCR) testing is considered the most accurate test available for COVID-19 diagnosis, it has some limitations, such as the need for specialized laboratory technicians and a long turnaround time. Therefore, we have established and reported a rapid diagnostic method using a small amount of saliva as a sample using a lightweight mobile qPCR device. This study aimed to improve the existing method and increase the detection sensitivity and specificity. The detection specificity of CDC N1 and N2 was examined by improving qPCR reagents and polymerase chain reaction conditions for the previously reported method. Furthermore, the feasibility of detecting severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) viral RNA was examined using both the previous method and the improved method in patients with COVID-19. The results showed that the improved method increased the specificity and sensitivity. This improved method is useful for the rapid diagnosis of SARS-CoV-2.

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Rapid and sensitive on-site detection of SARS-CoV-2 RNA from environmental surfaces using portable laboratory devices

Kitamura,

Ueno,

Yoshida

2023

Microbiol Spectr

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Monitoring severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) RNA on inanimate surfaces plays a crucial role in environmental surveillance, enabling the indirect detection of infected individuals and facilitating rapid infection control responses. However, developing a practical on-site, end-to-end method for detection remains a challenge. In this study, we established protocols for on-site detection of SARS-CoV-2 RNA on surfaces using two portable laboratory devices: The PicoGene PCR1100 and Bento Lab. Test particles comprised of inactivated viral particles containing partial sequences of SARS-CoV-2, type 1 poliovirus, coxsackie virus A16, and norovirus genogroups I and II were applied to reference materials as simulated contaminants. The direct detection method with the PicoGene PCR1100 achieved a recovery efficiency of 35.1% for the dried reference material. Furthermore, the Bento Lab, equipped with a microcentrifuge for on-site RNA purification, successfully detected a low concentration of inactivated SARS-CoV-2 reference standard. The recovery efficiency of the on-site RNA purification method on plastic, stainless steel, glass, and polyvinyl chloride surfaces ranged from 15.5% to 21.2%. Through the combination of these portable devices, SARS-CoV-2 RNA was detected within 1 h of swab sampling. This study contributes to the on-site risk assessment of infectious diseases by enabling the rapid detection of viral genomes. IMPORTANCE This study presents the development of a highly sensitive on-site method for detecting severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) RNA on various surfaces, including doorknobs and tables. Identifying SARS-CoV-2 RNA on these surfaces can be crucial in guiding decision-making for implementing non-pharmaceutical interventions, such as zoning strategies, improving ventilation, maintaining physical distancing, and promoting increased hand hygiene practices. Moreover, the on-site detection system can facilitate the swift initiation of mitigation responses in non-laboratory settings, including long-term care facilities and schools. The protocols established in this study offer a comprehensive approach for achieving both sensitivity and rapidity in on-site SARS-CoV-2 RNA detection. Furthermore, since the RT-qPCR assay serves as the gold standard for detecting viral RNAs, the developed protocol holds potential for application to other viruses, including enteroviruses and noroviruses.

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Feasibility of Rapid Diagnostic Technology for SARS-CoV-2 Virus Using a Trace Amount of Saliva

Cited by 3 publications

References 49 publications

Surface Functionalization of Non-Woven Fabrics Using a Novel Silica-Resin Coating Technology: Antiviral Treatment of Non-Woven Fabric Filters in Surgical Masks

Surface Functionalization of Non-Woven Fabrics Using a Novel Silica-Resin Coating Technology: Antiviral Treatment of Non-Woven Fabric Filters in Surgical Masks

Improving the Detection Sensitivity of a New Rapid Diagnostic Technology for Severe Acute Respiratory Syndrome Coronavirus 2 Using a Trace Amount of Saliva

Rapid and sensitive on-site detection of SARS-CoV-2 RNA from environmental surfaces using portable laboratory devices

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