基于CRISPR-Cas13a的GJB2c.235delC突变的快速检测,与侧向流量测量棒相结合
Yueqin Deng1, Juan Xu1, Ming Yang1
1Department of Otolaryngology Head and Neck Surgery, The Third Hospital of Changsha, Changsha 410015, Hunan, China.
Open life sciences
|March 17, 2025
概括
一个新的CRISPR-Cas核酸检测套件快速识别了遗传性听力损失的常见GJB2 c.235delC突变. 这种临床检测方法提供了快速,准确和可访问的诊断解决方案.
科学领域:
- 遗传学 遗传学 是一个
- 分子生物学分子生物学
- 生物技术是生物技术.
背景情况:
- 遗传性听力损失是一种流行的听力神经病变,对生活质量产生重大影响.
- 在中国人群中,GJB2 c.235delC突变是常见的原因.
- 现有的GJB2突变基因测试是昂贵的,耗时的,阻碍了临床应用.
研究的目的:
- 开发和评估用于GJB2 c.235delC突变的快速临床检测 (POCT) 工具包.
- 评估将CRISPR-Cas核酸检测与复合酶辅助放大 (RAA) 和侧流测量棒 (LFD) 集成到性基因查中的可行性.
主要方法:
- 集成的CRISPR-Cas13a核酸检测与RAA和LFD用于GJB2突变识别.
- 开发了一种能够检测低丰度核酸 (10^2副本/μL) 和低突变频率 (20%) 的POCT套件.
- 验证了来自31名聋患者和5名健康志愿者的血液样本的测定,证实了Sanger测序的结果.
主要成果:
- 在42分钟内成功识别了GJB2c.235delC突变.
- 该试验准确地区分了野生类型,同卵性和异卵性突变.
- 达到100%的诊断灵敏度和特异性,与桑格测序一致.
结论:
- CRISPR-Cas13a和LFD组合提供了一个快速而准确的方法来检测GJB2突变.
- 这种综合方法对与聋相关的基因的临床检测具有前景.
- 开发的POCT套件为可访问的遗传性听力损失诊断提供了可行的解决方案.
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