Highly accurate and sensitive diagnostic detection of SARS-CoV-2 by digital PCR

Li, Dong; Zhou, Jian; Niu, Chunyan; Wang, Quanyi; Pan, Yang; Sheng, Sitong; Wang, Xia; Zhang, Yongzhuo; Yang, Jiayi; Liu, Man‐Qing; Zhao, Yang; Zhang, Xiaoying; Zhu, Tao; Peng, Tao; Xie, Jun; Gao, Yunhua; Wang, Di; Dai, Xinhua; Fang, Xiang

doi:10.1016/j.talanta.2020.121726

Cited by 117 publications

(97 citation statements)

References 16 publications

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“…ddPCR is also more sensitive for detecting low target copies and relatively insensitive to potential PCR inhibitors (198). Recent studies have reported higher sensitivity and robustness of ddPCR than RT-PCR for detection and quantification of SARS-CoV-2 RNA from purified RNA and crude lysate samples in UTM (196,199). Digital droplet assays which enable detection and quantification with limited sample processing could potentially be used for monitoring clinical course and convalescence (199).…”

Section: Other Methods Of Viral Rna Detectionmentioning

confidence: 99%

Review of Current COVID-19 Diagnostics and Opportunities for Further Development

Mardian¹,

Kosasih²,

et al. 2021

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Diagnostic testing plays a critical role in addressing the coronavirus disease 2019 (COVID-19) pandemic, caused by Severe Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV-2). Rapid and accurate diagnostic tests are imperative for identifying and managing infected individuals, contact tracing, epidemiologic characterization, and public health decision making. Laboratory testing may be performed based on symptomatic presentation or for screening of asymptomatic people. Confirmation of SARS-CoV-2 infection is typically by nucleic acid amplification tests (NAAT), which requires specialized equipment and training and may be particularly challenging in resource-limited settings. NAAT may give false-negative results due to timing of sample collection relative to infection, improper sampling of respiratory specimens, inadequate preservation of samples, and technical limitations; false-positives may occur due to technical errors, particularly contamination during the manual real-time polymerase chain reaction (RT-PCR) process. Thus, clinical presentation, contact history and contemporary phyloepidemiology must be considered when interpreting results. Several sample-to-answer platforms, including high-throughput systems and Point of Care (PoC) assays, have been developed to increase testing capacity and decrease technical errors. Alternatives to RT-PCR assay, such as other RNA detection methods and antigen tests may be appropriate for certain situations, such as resource-limited settings. While sequencing is important to monitor on-going evolution of the SARS-CoV-2 genome, antibody assays are useful for epidemiologic purposes. The ever-expanding assortment of tests, with varying clinical utility, performance requirements, and limitations, merits comparative evaluation. We herein provide a comprehensive review of currently available COVID-19 diagnostics, exploring their pros and cons as well as appropriate indications. Strategies to further optimize safety, speed, and ease of SARS-CoV-2 testing without compromising accuracy are suggested. Access to scalable diagnostic tools and continued technologic advances, including machine learning and smartphone integration, will facilitate control of the current pandemic as well as preparedness for the next one.

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Section: Other Methods Of Viral Rna Detectionmentioning

confidence: 99%

Review of Current COVID-19 Diagnostics and Opportunities for Further Development

Mardian¹,

Kosasih²,

et al. 2021

View full text Add to dashboard Cite

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“…Four commercially available, one customized from Pasteur Institute and in-house optimized RT-PCR protocols (description available in Table 1 ) 6 , 7 were compared. Primer probes design, reaction mix and thermal cycling conditions are shown in Table 2 , 3 and 4 respectively.…”

Section: Methodsmentioning

confidence: 99%

“…No. 11732-088 Available upon request from Pasteur Institute In-house customized RT-PCR protocol Oncobit https://www.cdc.gov/coronavirus/2019-ncov/lab/rt-pcr-panel-primer-probes.html , September 7, 2020; 7 Ordered from Microsynth TaqPath 1-Step Multiplex Master Mix (no ROX) (A28521, ThermoFisher) SARS-CoV-2 Positive Run Control (#COV019CE) of Bio-Rad …”

Section: Methodsmentioning

confidence: 99%

A Comparative Study of Real-Time RT-PCR–Based SARS-CoV-2 Detection Methods and Its Application to Human-Derived and Surface Swabbed Material

Tastanova¹,

Stoffel²,

Dzung³

et al. 2021

The Journal of Molecular Diagnostics

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“…Moreover, dPCR showed higher sensitivity than RT‐qPCR in the testing of clinical samples. Some samples from patients with confirmed COVID‐19 were classified as positive by dPCR but found inconclusive or negative by RT‐qPCR 16,18–22 . For example, Suo et al 16 .…”

Section: Applications Of Digital Pcr In Covid‐19 Pandemicmentioning

confidence: 99%

Applications of digital PCR in COVID‐19 pandemic

Tan

Fan

Wang

et al. 2021

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The coronavirus disease 2019 (COVID‐19) pandemic has led to a public health crisis and global panic. This infectious disease is caused by a novel coronavirus named severe acute respiratory syndrome coronavirus 2 (SARS‐CoV‐2). Digital polymerase chain reaction (dPCR), which is an emerging nucleic acid amplification technology that allows absolute quantification of nucleic acids, plays an important role in the detection of SARS‐CoV‐2. In this review, we introduce the principle and advantages of dPCR, and review the applications of dPCR in the COVID‐19 pandemic, including detection of low copy number viruses, measurement of the viral load, preparation of reference materials, monitoring of virus concentration in the environment, detection of viral mutations, and evaluation of anti‐SARS‐CoV‐2 drugs. We also discuss the challenges of dPCR in clinical practice.

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Highly accurate and sensitive diagnostic detection of SARS-CoV-2 by digital PCR

Cited by 117 publications

References 16 publications

Review of Current COVID-19 Diagnostics and Opportunities for Further Development

Review of Current COVID-19 Diagnostics and Opportunities for Further Development

A Comparative Study of Real-Time RT-PCR–Based SARS-CoV-2 Detection Methods and Its Application to Human-Derived and Surface Swabbed Material

Applications of digital PCR in COVID‐19 pandemic

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