A one-step, one-tube real-time RT-PCR based assay with an automated analysis for detection of SARS-CoV-2

Dharavath, Bhasker; Yadav, Neelima; Desai, Sanket; Sunder, Roma; Mishra, Rohit; Ketkar, Madhura; Bhanshe, Prasanna; Gupta, Atul; Redhu, Archana Kumari; Patkar, Nikhil; Dutt, Shilpee; Gupta, Sudeep; Dutt, Amit

doi:10.1016/j.heliyon.2020.e04405

Cited by 35 publications

(37 citation statements)

References 28 publications

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“…In practice, diagnostic assays reliant on nucleic acid detection, especially Polymerase Chain Reaction (PCR)-based systems, are considered as the gold standard method. These assays include reverse transcription PCR (RT-PCR), real-time RT-PCR (rRT - PCR), and reverse transcription loop-mediated isothermal amplification (RT - LAMP), requiring specific primers and probes to SARS-CoV-2 genes, coding the first open reading frames [ 51 ], RNA-dependent RNA polymerase [ 52 ], envelope proteins [ 53 ], and nucleocapsids [ 54 ]. As faced during the pandemic, sampling errors, primer/probe designs, and RNA isolation are still impeding the assay performance and accuracy.…”

Section: Current Diagnostic Strategiesmentioning

confidence: 99%

Smart materials-integrated sensor technologies for COVID-19 diagnosis

et al. 2021

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After the first case has appeared in China, the COVID-19 pandemic continues to pose an omnipresent threat to global health, affecting more than 70 million patients and leading to around 1.6 million deaths. To implement rapid and effective clinical management, early diagnosis is the mainstay. Today, real-time reverse transcriptase (RT)-PCR test is the major diagnostic practice as a gold standard method for accurate diagnosis of this disease. On the other side, serological assays are easy to be implemented for the disease screening. Considering the limitations of today's tests including lengthy assay time, cost, the need for skilled personnel, and specialized infrastructure, both strategies, however, have impediments to be applied to the resource-scarce settings. Therefore, there is an urgent need to democratize all these practices to be applicable across the globe, specifically to the locations comprising of very limited infrastructure. In this regard, sensor systems have been utilized in clinical diagnostics largely, holding great potential to have pivotal roles as an alternative or complementary options to these current tests, providing crucial fashions such as being suitable for point-of-care settings, cost-effective, and having short turnover time. In particular, the integration of smart materials into sensor technologies leverages their analytical performances, including sensitivity, linear dynamic range, and specificity. Herein, we comprehensively review major smart materials such as nanomaterials, photosensitive materials, electrically sensitive materials, their integration with sensor platforms, and applications as wearable tools within the scope of the COVID-19 diagnosis.

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Section: Current Diagnostic Strategiesmentioning

confidence: 99%

Smart materials-integrated sensor technologies for COVID-19 diagnosis

et al. 2021

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“…It is therefore necessary to have an adequate knowledge of its reservoir, its distribution method and the best accessible management as well as preventive measures [31] , [32] , [33] . Closer monitoring and awareness remain a primary concern for doctors as well as health officials alike [34] , [35] . The community spread of the viral infection emphasizes the significance of the clinical diagnosis of illness caused by SARS-CoV-2, SARS-CoV and MERS-CoV.…”

Section: Introductionmentioning

confidence: 99%

SARS-CoV, MERS-CoV and SARS-CoV-2: A Diagnostic Challenge

2021

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Highlights The highly pathogenic MERS-CoV, SARS-CoV and SARS-CoV-2 cause acute illness and are often fatal. MERS-CoV, SARS-CoV and SARS-CoV-2 tend to infect the respiratory tract and can cause acute respiratory syndrome. SARS-related mortality rate is relatively higher than influenza. CT scans and RT-PCR are the most promising diagnostic tools for COVID-19 diagnosis.

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“…COVID-19 can be diagnosed in the laboratory by detecting SARS-CoV-2 genes in clinical samples collected from suspected patients, followed by viral isolation and culture [10]. Real-time RT-PCR is commonly used worldwide to diagnose COVID-19 in the laboratory setting [9,11,12].…”

Section: Introductionmentioning

confidence: 99%

Validation of Real-Time RT-PCR for Detection of SARS-CoV-2 in the Early Stages of the COVID-19 Outbreak Without Emergency Use Authorization Reagents in the Republic of Korea

Chung¹,

Lee²,

Woo³

et al. 2021

Preprint

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Background: A real-time reverse transcription polymerase chain reaction (RT-PCR) assay for detecting severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) was developed to rapidly diagnose coronavirus disease 2019 (COVID-19). Early diagnosis of COVID-19 enables timely treatment and the implementation of public health measures. We validated the sensitivity, specificity, precision, linearity, accuracy, and robustness of a real-time RT-PCR-based assay for SARS-CoV-2 detection and compared the sensitivity and specificity of Emergency Use Authorization (EUA)-approved reagents for COVID-19 with those of the real-time RT-PCR method.Methods: The real-time RT-PCR-based assay was performed to specifically amplify genomic markers of SARS-CoV-2.Results: The real-time RT-PCR assay was highly specific for SARS-CoV-2, and did not amplify genomic fragments of 13 other viruses that cause respiratory diseases. The assay showed high linearity when conducted with a viral isolate from a patient with COVID-19 together with plasmids containing the target genes. The assay showed good repeatability and reproducibility, with a coefficient of variation of 3%, and detected SARS-CoV-2 with a limit of detection of 1 PFU/mL.Conclusions: The present real-time RT-PCR-based assay can diagnose COVID-19 with high accuracy and sensitivity. This approach is highly effective and can facilitate the early diagnosis of COVID-19 without the use of EUA reagents in the Republic of Korea.

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A one-step, one-tube real-time RT-PCR based assay with an automated analysis for detection of SARS-CoV-2

Cited by 35 publications

References 28 publications

Smart materials-integrated sensor technologies for COVID-19 diagnosis

Smart materials-integrated sensor technologies for COVID-19 diagnosis

SARS-CoV, MERS-CoV and SARS-CoV-2: A Diagnostic Challenge

Validation of Real-Time RT-PCR for Detection of SARS-CoV-2 in the Early Stages of the COVID-19 Outbreak Without Emergency Use Authorization Reagents in the Republic of Korea

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