相关实验视频

Updated: Jun 25, 2025

Field-Deployable Candidatus Liberibacter asiaticus Detection Using Recombinase Polymerase Amplification Combined with CRISPR-Cas12a

Field-Deployable Candidatus Liberibacter asiaticus Detection Using Recombinase Polymerase Amplification Combined with CRISPR-Cas12a

Published on: December 23, 2022

使用Cas12f1核酶和工程指导RNA,有效,特定和直接检测双链DNA标

Jun He¹, Xipan Hu², Xingyong Weng³

¹The Affiliated Nanhua Hospital, Department of Clinical Laboratory, Hengyang Medical School, University of South China, Hengyang, China.

Biosensors & bioelectronics

|May 28, 2024

概括

一个新的基于CRISPR的生物传感器PDTCTR直接检测双链DNA (dsDNA) 以改善诊断. 这种先进的系统为检测传染病和癌症突变提供了显著更高的灵敏度.

科学领域:

在CRISPR诊断中使用CRISPR.
分子生物传感器分子生物传感器
核酸检测检测核酸检测的方法

背景情况:

CRISPR/Cas12f1系统需要复杂的ssDNA或RNA准备用于诊断.
现有的方法在灵敏度和直接dDNA检测方面存在局限性.

研究的目的:

开发一种基于CRISPR的新型生物传感器,用于直接检测双链DNA (dsDNA).
克服当前CRISPR诊断系统的局限性.

主要方法:

重组酶聚合酶放大 (RPA) 与 Cas12f_ge4.1 系统的集成.
开发了PAM-依赖的dsDNA目标激活的Cas12f1传输报告器 (PDTCTR) 生物传感器.
在没有先前核酸转化的情况下直接检测dDNA目标.

主要成果:

与最初的Cas12f1系统相比,PDTCTR的灵敏度提高了100倍.
在10个副本/μL时检测出Mycoplasma pneumoniae和B型肝炎病毒.
精确识别单核酸变异 (SNV),包括EGFR突变的NSCLC.
临床评估显示高灵敏度 (93%-100%) 和特异性 (100%).

结论:

PDTCTR可用于临床诊断的直接,高度敏感的dsDNA检测.
生物传感器显示出诊断传染病和与癌症相关的SNVs的巨大潜力.
克里斯普技术的进步为改善早期疾病检测和个性化医疗铺平了道路.

关键词:

(RPA) 的使用情况.Cas12f_ge4.1 系统的使用情况工程指导RNAs的工程指导RNAs.光生物传感器是一种光生物传感器.在PDTCTRTR中使用.复合酶聚合酶放大复合酶的放大.dsDNA检测检测的方法

更多相关视频

CIRCLE-Seq for Interrogation of Off-Target Gene Editing

CIRCLE-Seq for Interrogation of Off-Target Gene Editing

Published on: November 1, 2024

Enhanced Genome Editing with Cas9 Ribonucleoprotein in Diverse Cells and Organisms

Enhanced Genome Editing with Cas9 Ribonucleoprotein in Diverse Cells and Organisms

Published on: May 25, 2018

相关概念视频

CRISPR

CRISPR

Genome editing technologies allow scientists to modify an organism’s DNA via the addition, removal, or rearrangement of genetic material at specific genomic locations. These types of techniques could potentially be used to cure genetic disorders such as hemophilia and sickle cell anemia. One popular and widely used DNA-editing research tool that could lead to safe and effective cures for genetic disorders is the CRISPR-Cas9 system. CRISPR-Cas9 stands for Clustered Regularly Interspaced...

CRISPR and crRNAs

CRISPR and crRNAs

Bacteria and archaea are susceptible to viral infections just like eukaryotes; therefore, they have developed a unique adaptive immune system to protect themselves. Clustered regularly interspaced short palindromic repeats and CRISPR-associated proteins (CRISPR-Cas) are present in more than 45% of known bacteria and 90% of known archaea.
The CRISPR-Cas system stores a copy of foreign DNA in the host genome and uses it to identify the foreign DNA upon reinfection. CRISPR-Cas has three different...