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基于CRISPR-Cas13a系统的HIV-1感染高度敏感和快速的临床检测

Xiaohui Li^1,2, Bin Su¹, Lan Yang²

¹Beijing Key Laboratory for HIV/AIDS Research, Clinical and Research Center for Infectious Diseases, Beijing Youan Hospital, Capital Medical University, Beijing, 100069, China.

BMC infectious diseases

|September 25, 2023

概括

使用CRISPR-Cas13a和RT-RAA技术开发了一种针对人类免疫缺陷病毒第一类型 (HIV-1) RNA的新疗点检测 (POCT). 这种简单,敏感和特定的测试能够快速检测HIV-1RNA,用于早期诊断和治疗监测.

科学领域:

分子生物学分子生物学
病毒学病毒学
生物技术是生物技术.

背景情况:

人类免疫缺陷病毒第一型 (HIV-1) 导致获得性免疫缺陷综合征 (AIDS),这是一个全球性的健康问题.
抗逆转录病毒疗法抑制HIV-1,但需要终身坚持.
早期发现和诊断对于预防和控制艾滋病毒/艾滋病至关重要.
在高风险人群中对HIV-1进行管理,点诊所检测 (POCT) 是至关重要的.

研究的目的:

开发一种快速,准确和可访问的POCT用于HIV-1RNA检测.
使用CRISPR-Cas13a侧流带与逆转录酶复合酶辅助放大 (RT-RAA) 技术相结合.
为了使HIV-1RNA在护理点的视觉检测.

主要方法:

开发了一个POCT集成CRISPR-Cas13a侧流带与RT-RAA.
在RT-RAA初始设计中纳入了退化的基结CRISPR-Cas13a系统.
在临床样本上测试了用于检测HIV-1RNA的方法.

主要成果:

开发的POCT可以检测大多数HIV-1RNA菌株,检测极限 (LOD) 为1副本/μL.
没有与其他病原体显著的交叉反应.
在临床样本中实现了高性能指标:91.81%的灵敏度,100%的特异性,100%的正预测值 (PPV),39.14%的负预测值 (NPV) 和92.22%的准确性.
最低可检测的病毒载量为112个副本/毫升.

结论:

这种新的方法为HIV-1RNA提供了一个稳定,简单和高度敏感/特定的POCT.
这项技术具有很大的潜力,可以促进HIV-1感染的早期诊断.
POCT对于监测艾滋病毒患者治疗疗效非常有价值.

关键词:

这就是CRISPR-Cas13a.艾滋病病毒-1 艾滋病病毒-1 在护理场所进行检测.土耳其人土耳其人RT-RAAA

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