基于放大的CRISPR/Cas12a生物传感器针对COX1基因,用于特定检测猪DNA
Arifah A S Janudin1, Chitra P Kurup1, Lim Ya Chee2
1Biosensors and Nanobiotechnology Laboratory, Integrated Science Building, Faculty of Science, Universiti Brunei Darussalam, Jalan Tungku Link, Gadong BE1410, Brunei Darussalam.
ACS omega
|October 23, 2023
概括
一种新方法使用CRISPR/Cas12a和复合酶聚合酶放大 (RPA) 来检测食品中的猪肉. 这种敏感的测试提供了快速和具体的清真食品改的识别.
科学领域:
- 食品科学 食品科学 食品科学
- 生物技术是生物技术.
- 分子生物学分子生物学
背景情况:
- 使用未申报的猪肉成分改哈拉尔食品是全球范围内一个重大问题,特别是对于穆斯林消费者来说.
- 准确而敏感的检测方法对于确保食品完整性和消费者信任至关重要.
研究的目的:
- 开发一种快速,灵敏和特定的检测方法来检测食物样本中的猪DNA.
- 通过先进的分子技术,应对哈拉尔食品改的挑战.
主要方法:
- 开发了CRISPR/Cas12a介导的复合酶聚合酶放大 (RPA) 试验.
- 优化Cas12a裂变试剂度和指导RNA (gRNA) 选,以COX1基因为目标.
- 使用RPA和Cas12a联合切割系统检测猪DNA.
主要成果:
- 该试验成功检测到猪成分低至5 pg/μL,在线性范围5-1000 pg/μL内.
- 在没有预放大的情况下,实现了500倍低于基于CRISPR的方法的检测极限.
- 证明了高特异性,没有观察到与非目标物种DNA的交叉反应.
结论:
- 开发的CRISPR/Cas12a-RPA平台提供了一种高度敏感和特定的方法来检测猪DNA.
- 这种测试显示了快速选清真食品改的巨大潜力.
- 该方法为确保食品的真实性提供了一个有价值的工具.
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