使用CRISPR-Cas系统和侧向流量试验对乙型肝炎和C型肝炎病毒进行双重检测
Syeda Najidah Shahni1, Sarah Albogami2, Iqbal Azmi1
1Multidisciplinary Centre for Advanced Research and Studies, Jamia Millia Islamia, New Delhi, India.
Journal of nucleic acids
|October 14, 2024
概括
这项研究介绍了CRISPR-Cas12和CRISPR-Cas13用于敏感的乙型肝炎 (HBV) 和C型肝炎 (HCV) 病毒检测. 一种新的双酶侧流检测方法使得同时对这两种感染进行点诊断.
科学领域:
- 分子生物学分子生物学
- 生物技术是生物技术.
- 传染性疾病 传染性疾病
背景情况:
- 敏感和特定的诊断工具对于控制乙型肝炎病毒 (HBV) 和型肝炎病毒 (HCV) 至关重要.
- 当前的诊断方法在灵敏度和多重复合能力方面可能存在局限性.
- 克里斯普尔-卡斯系统为高度特定的核酸检测提供了潜力.
研究的目的:
- 开发和优化基于CRISPR-Cas12和CRISPR-Cas13的测定方法,用于HBV和HCV检测.
- 使用双酶方法提高诊断灵敏度和特异性.
- 创建一个多重侧流测定 (LFA) 用于同时检测HBV和HCV的护理点 (POC).
主要方法:
- 分别使用CRISPR-Cas12和CRISPR-Cas13系统检测HBV (DNA) 和HCV (RNA).
- 导向RNAs (gRNAs) 被设计和验证;通过凝电泳和光报告测试确认了裂变.
- 优化的gRNA被整合到横向流量试验 (LFA) 中,并实施了结合Cas12和Cas13的双酶策略.
主要成果:
- 优化的gRNAs通过LFA通过度依赖信号使HBV和HCV的敏感检测成为可能.
- 双酶方法显著改善了两种病毒的检测极限.
- 一个双抗原检测LFA条带成功地同时检测出HBV和HCV,而没有交叉反应.
结论:
- 克里斯普尔-卡斯系统可以有效地用于高度敏感和特定的HBV和HCV检测.
- 开发的双酶LFA为同时进行超敏感的POC诊断提供了一个有前途的平台.
- 这项技术解决了当前基于CRISPR的诊断技术的局限性,允许在单一测试条中进行多重检测.
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