Sensitive and Easy-Read CRISPR Strip for COVID-19 Rapid Point-of-Care Testing

Li, Hao; Dong, Xue; Wang, Yanhe; Yang, Lan; Cai, Kun; Zhang, Xiaolu; Kou, Zheng; He, Lihua; Sun, Shihui; Li, Tianyao; Nie, You; Li, Xiaofeng; Sun, Yuliang

doi:10.1089/crispr.2020.0138

Cited by 31 publications

(30 citation statements)

References 17 publications

(21 reference statements)

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“…The whole testing could be finished at room temperature by replacing PCR with isothermal amplification such as recombinase polymerase amplification ( 24 – 27 ). In most CRISPR-based assays, the readout is fluorescence signal, but the results could also be visualized by naked eyes, detected by mobile phone ( 32 ), or presented in a later flow strip ( 27 , 29 , 50 ). Therefore, it could be a rapid point-of-care assay for both diagnosis and epidemiologic surveillance of SARS-CoV-2 variants.…”

Section: Discussionmentioning

confidence: 99%

CRISPR-Cas12a-Based Detection for the Major SARS-CoV-2 Variants of Concern

et al. 2021

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We described CRISPR-Cas12-based multiplex allele-specific assay for rapid SARS-CoV-2 variant genotyping. The new system has the potential to be quickly developed, continuously updated, and easily implemented for screening of SARS-CoV-2 variants in resource-limited settings. This approach can be adapted for emerging mutations and implemented in laboratories already conducting SARS-CoV-2 nucleic acid amplification tests using existing resources and extracted nucleic acid.

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Section: Discussionmentioning

confidence: 99%

CRISPR-Cas12a-Based Detection for the Major SARS-CoV-2 Variants of Concern

et al. 2021

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“…Compared with previously reported CRISPR-based SARS-CoV-2 variant detection methods ( Zhang W. S. et al, 2021 ), ours provides a practical on-site test method to detect SARS-CoV-2 HV69-70del with obvious advantages. For example, strip-based PCR-CRISPR assay could eliminate the need for expensive fluorescence detectors, and improve the ability to diagnose SARS-CoV-2 variant on-site, particularly in under-developed and resource-limited regions ( Myers et al, 2013 ; Li H. et al, 2021 ). Besides, by combined with smartphone-assisted visualization tools ( Wen et al, 2021 ) or more accurate isothermal amplification, CRISPR detection system is expected to become a robust, rapid, quantitative and field-deployable POCT method for SARS-CoV-2 variants.…”

Section: Discussionmentioning

confidence: 99%

“…This experiment used test notes called ERASE lateral flow strips ( Li H. et al, 2021 ) that were previously developed by our research team. The ERASE strip was visualized and analyzed according to the “band-cutting method;” if, on the lateral flow strip, the C-band was visible, while the T-band was absent, a positive result was recorded, whereas if both the T-band and C-band were visible, the result was recorded as negative.…”

Section: Methodsmentioning

confidence: 99%

“…While, the limits of detection (LOD) and specificity of exist detection strategies are still need to improve for SARS-CoV-2 variants identification ( Patchsung et al, 2020 ). In this study, by combining our previously developed PCR-CRISPR genotypic detection ( Wang S. et al, 2021 ) and the easy-readout and sensitive enhanced (ERASE) lateral flow strip ( Li H. et al, 2021 ), we demonstrated a highly sensitive, and specific detection method for the SARS-CoV-2 HV69-70del. And this method facilitated us to identify SARS-CoV-2 variants in primary and resource limited medical institutions without professional and expensive fluorescent detector.…”

Section: Introductionmentioning

confidence: 99%

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Highly Sensitive Detection Method for HV69-70del in SARS-CoV-2 Alpha and Omicron Variants Based on CRISPR/Cas13a

Niu

Han

Dong

et al. 2022

Front. Bioeng. Biotechnol.

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As SARS-CoV-2 variants continue to evolve, identifying variants with adaptive diagnostic tool is critical to containing the ongoing COVID-19 pandemic. Herein, we establish a highly sensitive and portable on-site detection method for the HV69-70del which exist in SARS-CoV-2 Alpha and Omicron variants using a PCR-based CRISPR/Cas13a detection system (PCR-CRISPR). The specific crRNA (CRISPR RNA) targeting the HV69-70del is screened using the fluorescence-based CRISPR assay, and the sensitivity and specificity of this method are evaluated using diluted nucleic acids of SARS-CoV-2 variants and other pathogens. The results show that the PCR-CRISPR detection method can detect 1 copies/μL SARS-CoV-2 HV69-70del mutant RNA and identify 0.1% of mutant RNA in mixed samples, which is more sensitive than the RT-qPCR based commercial SARS-CoV-2 variants detection kits and sanger sequencing. And it has no cross reactivity with ten other pathogens nucleic acids. Additionally, by combined with our previously developed ERASE (Easy-Readout and Sensitive Enhanced) lateral flow strip suitable for CRISPR detection, we provide a novel diagnosis tool to identify SARS-CoV-2 variants in primary and resource-limited medical institutions without professional and expensive fluorescent detector.

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“…In this case, the authors considered the disappearance of the test band as positive, thus minimizing subjective readouts and improving the test accuracy. Validated with blind tests of 649 clinical samples, this assay achieved a test sensitivity of 90.67% and a specificity of 99.21% ( Li et al, 2021 ). Moreover, Fozouni and co-workers developed another Cas13a-based lateral flow strip for the detection of SARS-CoV-2 viral.…”

Section: Application Of Paper-based Poc Tests For Covid-19mentioning

confidence: 99%

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

Yuan

Sun

Tian

et al. 2021

Front. Bioeng. Biotechnol.

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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.

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Sensitive and Easy-Read CRISPR Strip for COVID-19 Rapid Point-of-Care Testing

Cited by 31 publications

References 17 publications

CRISPR-Cas12a-Based Detection for the Major SARS-CoV-2 Variants of Concern

CRISPR-Cas12a-Based Detection for the Major SARS-CoV-2 Variants of Concern

Highly Sensitive Detection Method for HV69-70del in SARS-CoV-2 Alpha and Omicron Variants Based on CRISPR/Cas13a

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

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