CRISPR/Cas12a-mediated ultrasensitive and on-site monkeypox viral testing

Zhao, Furong; Wang, Pei; Wang, Haoxuan; Liu, Sirui; Sohail, Muhammad; Zhang, Xing; Li, Bingzhi; Huang, He

doi:10.1039/d2ay01998a

Cited by 13 publications

(7 citation statements)

References 43 publications

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“…The lyophilized reagents can be stably stored at 4 °C for at least 4 weeks, making them suitable for on-site testing in resource-limited environments. The assay performances of isothermal amplication and CRISPR assays were summarized in Table S5, † 19,25,[29][30][31][32][33][34][35][36][37][38] which indicated the proposed single-step RPA-CRISPR/Cas12a assay exhibited a relative superior performance.…”

Section: Discussionmentioning

confidence: 99%

Detection of monkeypox virus based on a convenient and sensitive single-step RPA-CRISPR/Cas12a strategy

Yu,

Rong,

et al. 2024

RSC Adv.

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Section: Discussionmentioning

confidence: 99%

Detection of monkeypox virus based on a convenient and sensitive single-step RPA-CRISPR/Cas12a strategy

Yu,

Rong,

et al. 2024

RSC Adv.

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“…Until now, there are some monkeypox diagnostic methods that have been used for diagnosing the infections. Some of these techniques are polymerase chain reaction, , virus culture-based diagnosis, protein Cas-based diagnosis, , loop-mediated isothermal amplification, , enzyme-linked immunosorbent assay, , and recombinase polymerase amplification assay. ,, Considering the point-of-care (POC) nature of rapid tests, antibody-based immunoassays seem to be stronger diagnostic tool candidates for MPXV detection. However, due to the antigen context of Orthopoxvirus genomes, most human samples become cross-reactive, and this situation makes it impossible to differentiate different kinds of Orthopoxvirus viruses. − …”

Section: Introductionmentioning

confidence: 99%

Electrochemical Immunoassay Platform for Human Monkeypox Virus Detection

Can,

Perk,

Çitil

et al. 2024

Anal. Chem.

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In this study, we reported a selective impedimetric biosensor for the detection of A29 which is the target protein of the monkeypox virus (MPXV). The working principle of the biosensor relies on the interaction mechanism between A29, which is an internal membrane protein of MPXV, and the heparan sulfate receptor. For this purpose, after immobilizing heparan sulfate onto the gold screen-printed electrode surface, its interaction with A29 protein was monitored using electrochemical impedance spectroscopy. After the optimization of experimental parameters, the analytical characteristics of the developed MPVX immunosensor were examined. The developed immunosensor exhibited a linear detection range between 2.0 and 50 ng mL −1 , with a detection limit of 2.08 ng mL −1 and a quantification limit of 6.28 ng mL −1 . Furthermore, a relative standard deviation value of 2.82% was determined for 25 ng mL −1 . Apart from that, sample application studies were also performed with the standard addition of A29 protein to 1:10 diluted real serum samples that were taken from healthy individuals, and very good recovery values were obtained.

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“…CRISPR technology has been incorporated into the detection of nucleic acid, possessing excellent simplicity, specificity, and sensitivity that surpasses conventional assays. − The Cas12a nuclease recognizes DNA and gets activated, thereby trans -cleaving nearby single-stranded DNA . This unique activity has been employed for the sensitive and precise in vitro detection of virus genomes, including SHERLOCK, DETECTOR, and HOLMES, ,, providing potent diagnostic toolboxes for point-of-care testing (POCT). , Aiming to control the COVID-19 pandemic, several CRISPR-based POCT targeting SARS-CoV-2 RNA was developed, − revealing high potential in-home tests.…”

Section: Introductionmentioning

confidence: 99%

Active CRISPR-Cas12a on Hydrophilic Metal–Organic Frameworks: A Nanobiocomposite with High Stability and Activity for Nucleic Acid Detection

Zeng

Shang

et al. 2023

Anal. Chem.

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CRISPR-Cas12a is an accurate and responsive biosensing technique, but its limited stability has restricted its widespread applications. To address this, we propose a strategy using metal−organic frameworks (MOFs) to protect Cas12a from harsh environments. After screening multiple candidate MOFs, it was found that hydrophilic MAF-7 is highly compatible with Cas12a, and the as-formed Cas12a-on-MAF-7 (COM) not only retains high enzymatic activity but also possesses excellent tolerance to heat, salt, and organic solvents. Further investigation showed that COM can serve as an analytical component for nucleic acid detection, resulting in an ultrasensitive assay for SARS-CoV-2 RNA detection with a detection limit of 1 copy. This is the first successful attempt to create an active Cas12a nanobiocomposite that functions as a biosensor without the need for shell deconstruction or enzyme release.

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CRISPR/Cas12a-mediated ultrasensitive and on-site monkeypox viral testing

Abstract: The unexpected spread of the monkeypox virus (MPXV) from Central and West Africa to previously non-endemic regions has caused a global panic. In this context, the rapid, specific, and ultrasensitive...

Cited by 13 publications

References 43 publications

Detection of monkeypox virus based on a convenient and sensitive single-step RPA-CRISPR/Cas12a strategy

Detection of monkeypox virus based on a convenient and sensitive single-step RPA-CRISPR/Cas12a strategy

Electrochemical Immunoassay Platform for Human Monkeypox Virus Detection

Active CRISPR-Cas12a on Hydrophilic Metal–Organic Frameworks: A Nanobiocomposite with High Stability and Activity for Nucleic Acid Detection

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