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一个CRISPR/cas13a辅助的精确和便携式测试用于布鲁塞拉核酸检测

Haiwen Liu^1,2, Ling Xu³, Ying Xiu⁴

¹National Key Laboratory of Intelligent Tracing and Forecasting for Infectious Diseases, National Institute for Communicable Disease Control and Prevention, Chinese Center for Disease Control and Prevention, Beijing, China.

Frontiers in cellular and infection microbiology

|April 7, 2025

概括

这项研究引入了一种快速RAA-CRISPR/Cas13a检测布鲁塞拉细菌的方法. 这种核酸测试在各种样本类型中提供了一种敏感和特定的工具,用于早期诊断乳病.

科学领域:

兽医微生物学兽医微生物学
分子诊断学分子诊断
公共卫生公共卫生

背景情况:

由布鲁塞拉菌感染引起的菌病带来了重大的经济和公共卫生风险.
目前布鲁塞洛症的分子诊断方法通常很复杂,缺乏灵敏度,并且很难在现场应用.
对于快速,方便和现场诊断工具来检测布鲁塞拉有着至关重要的需求.

研究的目的:

开发一种快速方便的基于核酸的分子方法,用于现场检测布鲁塞拉菌.
建立一种早期临床查方法来检测血病.
在现实世界样本类型中验证开发方法的实际实用性.

主要方法:

基于保存的布鲁塞拉Bcsp31基因设计的CRISPRRNA (crRNA) 和RAA原料.
综合复合酶辅助放大 (RAA) 具有CRISPR/Cas13a检测.
开发了Brucella核酸的光和基于纸条的检测格式.

主要成果:

该RAA-CRISPR/Cas13a光方法实现了10^0副本/μL的检测极限,而条纹测试的极限为10^1副本/μL.
这两种方法在检测布鲁塞拉菌中都显示出100%的特异性.
牛奶,人体全血和绵羊阴道拭片的阳性率分别为93%,82.12%和91%的光,以及87%,64.23%和76%的条纹测试.

结论:

成功开发了一种针对布鲁塞拉BCSP31基因的新型RAA-CRISPR检测方法.
这种方法显示了Brucella核酸准确识别的巨大潜力.
开发的试验对临床诊断和brucellosis的现场查具有重要意义.

关键词:

布鲁塞拉病毒1 这就是CRISPR-Cas13a3的特征.在 RAA6A6 中.葡萄球菌病2 葡萄球病2 光检测4 光检测核酸检测5核酸检测5 条纹试验7 条纹试验7 动物病毒8 病毒8

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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...