Liu Peng-Fei, Zhao Kai-Ren, Liu Zhi-Jun, Wang Li, Ye Shu-Ying, Liang Guo-Xi
School of Pharmacy, Jiangsu University, Zhenjiang, 212013, China.
School of Pharmacy, Jiangsu University, Zhenjiang, 212013, China.
Biosens Bioelectron. 2021 Mar 15;176:112954. doi: 10.1016/j.bios.2020.112954. Epub 2021 Jan 1.
CRISPR/Cas system have drawn increasing attention in accurate and sensitive nucleic acids detection. Herein, we reported a novel Cas12a-based electrochemiluminescence biosensor for target amplification-free human papilloma virus subtype (HPV-16) DNA detection. During this detection process, Cas12a employed its two-part recognition mechanism to improve the specificity and trans-cleavage capability to achieve signal amplification, while L-Methionine stabilized gold nanoclusters (Met-AuNCs) were served as high-efficiency ECL emitters to achieve ECL signal transition. Given the unique combination of Cas12a with ECL technique, the detection limit was determined as 0.48 pM and the whole detection could be completed within 70 min. We also validated the practical application of the proposed biosensor by using undiluted human blood samples, which gives impetus to the design of new generations of CRISPR/Cas detection system beyond the traditional ones with ultimate applications in sensing analysis and diagnostic technologies.
CRISPR/Cas系统在准确灵敏的核酸检测中受到越来越多的关注。在此，我们报道了一种基于新型Cas12a的电化学发光生物传感器，用于无目标扩增的人乳头瘤病毒亚型（HPV-16）DNA检测。在该检测过程中，Cas12a利用其双识别机制提高特异性和反式切割能力以实现信号放大，同时L-甲硫氨酸稳定的金纳米簇（Met-AuNCs）作为高效电化学发光发射体实现电化学发光信号转换。鉴于Cas12a与电化学发光技术的独特结合，检测限确定为0.48 pM，整个检测可在70分钟内完成。我们还通过使用未稀释的人血样本验证了所提出生物传感器的实际应用，这推动了新一代CRISPR/Cas检测系统的设计，超越了传统系统，在传感分析和诊断技术中具有最终应用。
