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系统在准确和灵敏的核酸检测中引起了越来越多的关注。在这里，我们报告了一种新型的基于Cas12a的电化学发光生物传感器，用于无靶标扩增的人乳头瘤病毒亚型（HPV-16）DNA检测。在此检测过程中，Cas12a利用其两部分识别机制来提高特异性和反式裂解能力以实现信号放大，而L-蛋氨酸稳定的金纳米团簇（Met-AuNCs）被用作高效ECL发射体以实现ECL信号转换。鉴于Cas12a与ECL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类血液样本验证了所提出的生物传感器的实际应用，