对CRISPR/Cas系统稳定性的基本评估,用于基于纸张的分析设备.
Yohei Tanifuji1, Hikaru Suzuki1, Guodong Tong1
1Department of Applied Chemistry, Faculty of Science and Technology, Keio University, 3-14-1 Hiyoshi, Kohoku-ku, Yokohama, 223-8522, Japan. citterio@applc.keio.ac.jp.
Analytical methods : advancing methods and applications
|June 12, 2024
概括
在纸上干燥的CRISPR-Cas试剂在-20°C下28天后保持超过70%的活性,而糖糖提高了稳定性. 干燥试剂表现出增强的耐热性,使得简化,集成的点的护理诊断设备.
科学领域:
- 生物技术是生物技术.
- 分析化学 分析化学
- 分子诊断学 分子诊断
背景情况:
- 克里斯普尔/卡斯系统为临床核酸测试提供了潜力.
- 集成到基于纸张的分析设备 (PADs) 是有限的,纸张通常只用于读出.
- 基于CRISPR的纸质试验的设备上的试剂存储稳定性在很大程度上仍未被探索.
研究的目的:
- 为了研究CRISPR/Cas试剂在纸质基板上的存储稳定性.
- 评估影响试剂稳定性的因素,包括纸张类型,干燥方法,稳定剂和储存条件.
- 评估在设备上储存试剂的可行性,以进行简化,综合诊断分析.
主要方法:
- 在各种纸质基板上干燥CRISPR-Cas12a试剂.
- 通过测量ssDNA探针的跨裂变的光度来评估试剂活性.
- 在不同条件下在-20°C下保存28天后评估稳定性,包括使用和不使用诸如糖糖等稳定剂.
主要成果:
- 在BSA阻塞纸上干燥的CRISPR试剂在28天后在-20°C下保持超过70%的初始活性.
- 作为稳定剂添加的糖显著改善了试剂储存的稳定性.
- 与液体溶液相比,干燥试剂的热稳定性在65°C以上得到了提高.
结论:
- 基于CRISPR的测试试剂可以有效地存储在纸质基板上,简化了测试程序.
- 优化的干燥和稳定方法提高了试剂的寿命和纸张的耐热性.
- 这项研究支持开发完全集成的,试剂稳定的基于纸张的分析设备,用于临床诊断.
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