CRISPR-Based Assays for Rapid Detection of SARS-CoV-2

Javalkote, Vivek S.; Kancharla, Nagesh; Bhadra, Bhaskar; Shukla, Manish; Soni, Badrish; Sapre, Ajit; Goodin, Michael M.; Bandyopadhyay, Anindya; Dasgupta, Santanu

doi:10.20944/preprints202006.0025.v1

Cited by 11 publications

(7 citation statements)

References 25 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Currently, nucleic acid amplification tests (NAAT) are considered the recommended, most sensitive tests for SARS-CoV-2 detection ( Böger et al., 2021 ; Kevadiya et al., 2021 ). qRT-PCR is the gold standard molecular test ( Corman et al., 2020 ), and different targets at the ORF1ab, S, E, or N gene have been reported for molecular detection ( Broughton et al., 2020 ; Esbin et al., 2020 ; Huang et al., 2020 ; Javalkote et al., 2020 ). Differences in specificity, amplification efficiency, and also downstream detection (i.e., CRISPR-Cas) have been described ( Joung et al., 2020 ; Xiong et al., 2020 ).…”

Section: Resultsmentioning

confidence: 99%

Unlocking SARS-CoV-2 detection in low- and middle-income countries

Alcántara¹,

Peñaranda²,

Mendoza-Rojas³

et al. 2021

Cell Reports Methods

View full text Add to dashboard Cite

Low- and middle-income countries (LMICs) are significantly affected by SARS-CoV-2, partially due to their limited capacity for local production and implementation of molecular testing. Here, we provide detailed methods and validation of a molecular toolkit that can be readily produced and deployed using laboratory equipment available in LMICs. Our results show that lab-scale production of enzymes and nucleic acids can supply over 50,000 tests per production batch. The optimized one-step RT-PCR coupled to CRISPR-Cas12a-mediated detection showed a limit of detection of 10 2 ge/μL in a turnaround time of 2 h. The clinical validation indicated an overall sensitivity of 80%–88%, while for middle and high viral load samples (Cq ≤ 31) the sensitivity was 92%–100%. The specificity was 96%–100% regardless of viral load. Furthermore, we show that the toolkit can be used with the mobile laboratory Bento Lab, potentially enabling LMICs to implement detection services in unattended remote regions.

show abstract

Section: Resultsmentioning

confidence: 99%

Unlocking SARS-CoV-2 detection in low- and middle-income countries

Alcántara¹,

Peñaranda²,

Mendoza-Rojas³

et al. 2021

Cell Reports Methods

View full text Add to dashboard Cite

show abstract

“…Adapted with permission from ( Li et al, 2020 ) under Creative Commons license; (C) principle of a CRISPR test. Adapted with permission from ( Javalkote et al, 2020 ) under Creative Commons license.…”

Section: Paper-based Sars-cov-2 Diagnostic Methodsmentioning

confidence: 99%

Paper-Based Point-of-Care Testing of SARS-CoV-2

Yuan

Sun

Tian

et al. 2021

Front. Bioeng. Biotechnol.

View full text Add to dashboard Cite

The COVID-19 pandemic has resulted in significant global social and economic disruption. The highly transmissive nature of the disease makes rapid and reliable detection critically important. Point-of-care (POC) tests involve performing diagnostic tests outside of a laboratory that produce a rapid and reliable result. It therefore allows the diagnostics of diseases at or near the patient site. Paper-based POC tests have been gaining interest in recent years as they allow rapid, low-cost detection without the need for external instruments. In this review, we focus on the development of paper-based POC devices for the detection of SARS-CoV-2. The review first introduces the principles of detection methods that are available to paper-based devices. It then summarizes the state-of-the-art paper devices and their analytical performances. The advantages and drawbacks among methods are also discussed. Finally, limitations of the existing devices are discussed, and prospects are given with the hope to identify research opportunities and directions in the field. We hope this review will be helpful for researchers to develop a clinically useful and economically efficient paper-based platform that can be used for rapid, accurate on-site diagnosis to aid in identifying acute infections and eventually contain the COVID-19 pandemic.

show abstract

“…those concerning school populations or mass testing of large groups). 30,43,105,106 Self-sampling methods that provide increased sensitivity could broaden the capacity for mass testing prior to events or entering the workplace. 27 As it will take time before vaccination will be able to eradicate COVID-19, testing measures will continue to be important.…”

Section: Vaccinated Individualsmentioning

confidence: 99%