Biosensing platforms for DNA diagnostics based on CRISPR/Cas nucleases: towards the detection of nucleic acids at the level of single molecules in non-laboratory settings DOI
Svetlana A. Khmeleva, K. G. Ptitsyn, Leonid K. Kurbatov

et al.

Biomeditsinskaya Khimiya, Journal Year: 2024, Volume and Issue: 70(5), P. 287 - 303

Published: Jan. 1, 2024

The use of CRISPR/Cas nucleases for the development DNA diagnostic systems in out-of-laboratory conditions (point-of-need testing, PONT) has demonstrated rapid growth last few years, starting with appearance 2017–2018 first platforms known as DETECTR and SHERLOCK. are based on a combination methods nucleic acid isothermal amplification selective detection target amplicons. This significantly improves sensitivity specificity PONT, making them comparable or even superior to polymerase chain reaction, considered “gold standard” diagnostics. review considers modern approaches coupling using Cas9, Cas12a, Cas12b, Cas13a, Cas14, Cas3 various amplification, an emphasis works which at level single molecules (attomolar subattomolar concentrations target) is achieved. properties used targeted diagnostics features briefly context biosensing platforms. Special attention paid most promising directions nuclease.

Language: Английский

Field-Deployable Detection of Genetically Modified Organisms with an Integrated Method of Loop-Mediated Isothermal Amplification and CRISPR/FnCas12a DOI
Lu Chen, Chen Wang,

Zaobing Zhu

et al.

Journal of Agricultural and Food Chemistry, Journal Year: 2025, Volume and Issue: 73(9), P. 5625 - 5634

Published: Feb. 18, 2025

The detection of genetically modified organisms (GMOs) is crucial for regulatory compliance and consumer safety. This study presents a novel method combining loop-mediated isothermal amplification (LAMP) with CRISPR/Cas12a cleavage, termed Cas-pfLAMP, sensitive specific GMO detection. We developed assays three GM events: maize DBN9936 MON810 soybean GTS40-3-2. By incorporating universal protospacer adjacent motif (PAM) sequence into LAMP primers, we overcame the limitations PAM site dependence. Cas-pfLAMP demonstrated high specificity sensitivity, limits as low 10-12 copies per reaction. Furthermore, point-of-care testing platform integrating rapid DNA extraction, lateral flow strips on-site achieved comparable sensitivity to qPCR, detecting contents 0.1% in simulated samples within 40 min. offers advantages independence, suitability field without specialized equipment. approach represents promising new generation methods potential applications various scenarios.

Language: Английский

Citations

0
Ultrasensitive and visual identification of lethal mushroom Russula subnigricans by LAMP-based CRISPR/Cas12a assay DOI
Zhiyong Zhao,

Lanxin Zhao,

Enjing Tian

et al.

Microchemical Journal, Journal Year: 2025, Volume and Issue: unknown, P. 113100 - 113100

Published: Feb. 1, 2025

Language: Английский

Citations

0
Viral Contamination of Food DOI
Long Ma,

Weipan Peng,

Junjie Wang

et al.

Elsevier eBooks, Journal Year: 2025, Volume and Issue: unknown, P. 79 - 92

Published: Jan. 1, 2025

Language: Английский

Citations

0
An extraction-free one-step pathogen detection system (Ex-opods) based on LAMP and CRISPR-Cas12b DOI

Wenjing Ren,

Weixin You, Hongmei Xu

et al.

Microchemical Journal, Journal Year: 2025, Volume and Issue: unknown, P. 113724 - 113724

Published: April 1, 2025

Language: Английский

Citations

0
Biosensing platforms for DNA diagnostics based on CRISPR/Cas nucleases: towards the detection of nucleic acids at the level of single molecules in non-laboratory settings DOI
Svetlana A. Khmeleva, K. G. Ptitsyn, Leonid K. Kurbatov

et al.

Biomeditsinskaya Khimiya, Journal Year: 2024, Volume and Issue: 70(5), P. 287 - 303

Published: Jan. 1, 2024

The use of CRISPR/Cas nucleases for the development DNA diagnostic systems in out-of-laboratory conditions (point-of-need testing, PONT) has demonstrated rapid growth last few years, starting with appearance 2017–2018 first platforms known as DETECTR and SHERLOCK. are based on a combination methods nucleic acid isothermal amplification selective detection target amplicons. This significantly improves sensitivity specificity PONT, making them comparable or even superior to polymerase chain reaction, considered “gold standard” diagnostics. review considers modern approaches coupling using Cas9, Cas12a, Cas12b, Cas13a, Cas14, Cas3 various amplification, an emphasis works which at level single molecules (attomolar subattomolar concentrations target) is achieved. properties used targeted diagnostics features briefly context biosensing platforms. Special attention paid most promising directions nuclease.

Language: Английский

Citations

0