通过crRNA工程设计的CRISPR/Cas12a系统与RPA相结合,用于超灵敏地检测Lactiplantibacillus plantarum
Ying Yu1, Shuxia Sun1, Xin Song1
1Shanghai Engineering Research Center of Food Microbiology, School of Health Science and Engineering, University of Shanghai for Science and Technology, Shanghai 200093, China.
Food chemistry
|February 28, 2026
概括
一个新的RPA-ECas12a平台可以快速检测Lactiplantibacillus plantarum益生菌. 这种增强的CRISPR/Cas12a系统为食品工业提供了精确而高效的细菌菌株识别.
科学领域:
- 微生物学 微生物学
- 分子生物学分子生物学
- 生物技术是生物技术.
背景情况:
- 不断扩大的益生菌市场需要先进的方法来准确识别细菌菌株.
- 目前的检测技术可能耗时,缺乏强大的质量控制所需的灵敏度.
研究的目的:
- 开发一种快速,灵敏和精确的单式光平台,用于检测Lactiplantibacillus plantarum.
- 设计一个CRISPR/Cas12a系统,以提高诊断性能.
主要方法:
- 重组酶聚合酶放大 (RPA) 与增强的CRISPR/Cas12a系统 (RPA-ECas12a) 的整合.
- 合理的CRISPRRNA (crRNA) 的5端DNA延伸以优化Cas12a跨裂变活性,确定一个最佳变体 (5'crRNA10).
- 该平台应用于复杂的食品矩阵,包括益生菌粉末和酸奶.
主要成果:
- 优化的5'crRNA10变体提高了Cas12a的催化效率,与野生类型crRNA相比提高了33%.
- 对于L. plantarum,RPA-ECas12a平台实现了1.3 CFU/mL的检测极限.
- 检测在45分钟内完成,精度高 (CVs<10%),在各种食品样本中具有选择性.
结论:
- 开发的RPA-ECas12a平台为益生菌认证提供了一个灵敏,快速和强大的工具.
- 在诊断应用中,crRNA工程策略可用于提高CRISPR/Cas12a性能.
- 这种方法支持不断增长的益生菌行业的质量控制和安全.
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