基于CRISPR/Cas12a的检测平台,用于早期和快速诊断草皮伤寒
Pooja Bhardwaj1, Nikita Shrikant Nanaware2, Sthita Pragnya Behera1
1ICMR-Regional Medical Research Centre Gorakhpur, BRD Medical College Campus, Gorakhpur 273013, India.
Biosensors
|December 22, 2023
概括
一种新的基于CRISPR/Cas12a的横向流量测定 (LoCIST) 提供快速,灵敏和无仪器检测灌木伤寒 (ST). 这种临床诊断工具有助于控制急性脑炎综合征爆发.
科学领域:
- 分子生物学分子生物学
- 传染病 传染病 传染病
- 诊断 诊断 诊断 诊断
背景情况:
- 由Orientia tsutsugamushi引起的草皮伤寒 (ST) 是急性脑炎综合征 (AES) 的重要原因.
- 及时及现场检测ST对于有效控制AES爆发至关重要.
研究的目的:
- 开发一种快速,灵敏和无仪器的诊断方法来诊断灌木伤寒.
- 为了评估基于CRISPR/Cas12a的新型侧向流量试验 (LoCIST) 对于ST检测的性能.
主要方法:
- 开发一个三步测定方法:同热放大,CRISPR/Cas12a识别和侧流测定 (LFA) 检测.
- 利用复合酶聚合酶用于异热放大和CRISPR/Cas12a技术用于目标识别.
主要成果:
- 通过LoCIST测定,每个反应达到一个基因拷贝的检测极限,并在一个小时内完成检测.
- 在81个临床样本上进行测试,LoCIST显示97.6%的灵敏度和100%的特异性,没有与其他瑞克特 DNA 交叉反应.
- 测试结果与ST的PCR检测100%一致.
结论:
- LoCIST提供了一种新的,便携式的,简单的,敏感的和特定的方法来检测灌木伤寒.
- 由于LoCIST的无仪器性质,它非常适合在护理点进行诊断和控制AES爆发.
相关概念视频
CRISPR and crRNAs
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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...
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