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为了Chlamydia psittaci的检测,LAMP-CRISPR/Cas12b平台的工程

Rong Wang¹, Xujian Mao², Jian Xu²

¹School of Public Health, Nanjing Medical University, Nanjing, Jiangsu 211166, PR China.

Journal of medical microbiology

|December 6, 2023

概括

一种新的LAMP-CRISPR/Cas12b测定方法快速检测出克拉米迪亚虫 (C. psittaci),这是导致虫病的原因. 这种敏感和特定的方法为这种动物传播感染提供了快速诊断.

科学领域:

分子生物学分子生物学
传染性疾病传染性疾病
生物技术是生物技术.

背景情况:

克拉米迪亚皮斯塔奇 (C. psittaci) 是一种致动物性细菌,在人类中引起皮斯塔科斯 (热).
严重的临床表现包括肺炎,ARDS和潜在的死亡,需要快速诊断.
目前的检测方法可能缺乏及时干预所需的速度,灵敏度或特异性.

研究的目的:

设计一个新的LAMP-CRISPR/Cas12b平台,用于快速检测C. psittaci.
建立双步和单管反应系统,以提高诊断能力.

主要方法:

结合循环介导的同热放大 (LAMP) 与CRISPR-Cas12b测定.
设计了特定的LAMP原料和sgRNA,针对C. psittaci的保存的CPSIT_0429基因.
与qPCR相比,对临床样本进行评估检测极限,特异性和性能.

主要成果:

开发了两步和一管LAMP-CRISPR/Cas12b测定,可以在1小时内检测.
证明了高特异性,没有与非C. psittaci模板的交叉反应.
在160个临床样本中达到10^2 aM (两步) 和10^3 aM (一管) 的低检测极限,结果与qPCR一致.

结论:

开发的LAMP-CRISPR/Cas12b测定是一种敏感和特定的工具,用于快速检测C. psittaci.
这个平台为及时诊断松病提供了至关重要的技术支持.
提供了一个有前途的进步分子诊断动物性细菌感染的分子诊断.

关键词:

这就是CRISPR-Cas12b.克拉米迪亚皮斯塔西西一个灯的灯光.检测检测检测检测检测

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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...