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在CRISPR-Cas12a的基础上检测和区分Mycobacterium sppp

Peeraphan Compiro¹, Nantinee Chomta¹, Juthamas Nimnual¹

¹Center of Excellence in Systems Microbiology (CESM), Department of Biochemistry, Faculty of Medicine, Chulalongkorn University, Rama IV, Bangkok 10330, Thailand.

Clinica chimica acta; international journal of clinical chemistry

|December 26, 2024

概括

一种新的MyTRACK测定方法快速检测和区分使用异热RPA和CRISPR-Cas12a.a的Mycobacterium物种. 这种精确的诊断工具非常适合识别结核病和其他真菌细菌感染,特别是在资源有限的环境中.

科学领域:

微生物学微生物学
分子诊断学分子诊断
传染性疾病传染性疾病

背景情况:

菌根杆菌的种类导致人类的重大疾病,如结核病.
准确和快速的诊断对于有效的治疗和疾病控制至关重要.
目前的诊断方法可能缺乏速度或特异性来区分菌根菌种.

研究的目的:

开发一种新的异热测定方法,MyTRACK,用于同时检测和分化主要临床菌根菌的物种水平.
将复合酶聚合酶放大 (RPA) 和CRISPR-Cas12a技术结合起来,以提高诊断能力.

主要方法:

开发一种结合RPA和CRISPR-Cas12a的异热分析方法.
使用已知的真菌细菌DNA测试试验的特异性和检测极限 (LOD).
在临床分离物和样本上验证MyTRACK试验,将结果与培养物和线索探测试验 (LPA) 进行比较.

主要成果:

MyTRACK试验表明没有交叉反应性和低检测极限 (1-100副本/反应) 的向菌根.
通过临床分离物实现了100%的特异性和92.59%至100%的敏感性.
临床样本的结果与传统培养和LPA方法一致.

结论:

MyTRACK测定是一种高效,便携式,快速和准确的菌根细菌检测和识别方法.
这种测试显示出在缺乏资源的临床环境中使用的巨大潜力,改善了结核病和非结核性菌根感染的诊断.

关键词:

这就是CRISPR-Cas12a.检测检测检测检测检测这是Mycobacterium.在 RPA RPA 中使用.结核病是一种疾病.rpoBB 的时间.

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相关概念视频

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...