Recent advances in CRISPR/Cas biosensing have led to impressive performance in sensitivity, specificity, and speed for nucleic acid detection. However, the remarkable advantages (such as universality, ultralow, attomolar detection limits) of CRISPR/Cas biosensing systems are limited in testing non-nucleic acid targets. Herein, by synthesizing a functional hybrid conjugate of antibody and single strand DNA oligonucleotide, we had successfully demonstrated the capability to integrate CRISPR/Cas12a-based signal amplification into different types of immunoassay schemes without the need for any additional recognition molecule or molecular synthesis during the detection process, thus providing a simple but generally applicable approach to improve the conventional immunoassays with attomolar sensitivity for small protein detections, referred as the CRISPR-based Universal Immunoassay Signal Enhancer (CRUISE). CRUISE is capable of being integrated into various immunoassays either through the primary antibody or the secondary antibody, with sensitivity down to 1 fg mL⁻¹ (∼50 aM) and 6 logs of linear range for detecting cytokines, such as IFN-γ and EGFR, under 3–4 h. It has a 10³ times higher sensitivity compared to a commercial IFN-γ ELISA kit, but uses the same experimental scheme. The same 1 fg mL⁻¹ sensitivity along with 6 logs of linear range was realized for IFN-γ detection in human plasma samples. We are expecting that our CRUISE provides an alternative but simple, user-friendly and effective strategy for those who rely on the use of immunoassays, while struggling with the limits of their sensitivity or detection ranges.

CRISPR/Cas 生物传感的最新进展已在核酸检测的灵敏度、特异性和速度方面取得了令人瞩目的表现。然而，CRISPR/Cas 生物传感系统的显着优势（如通用性、超低、阿摩尔级检测限）在检测非核酸靶点时受到限制。在此，通过合成抗体和单链 DNA 寡核苷酸的功能性杂交偶联物，我们成功地证明了将基于 CRISPR/Cas12a 的信号放大整合到不同类型的免疫测定方案中的能力，而无需任何额外的识别分子或分子合成。检测过程，从而提供了一种简单但普遍适用的方法来改进常规免疫测定法，对小蛋白质检测具有阿摩尔级灵敏度，称为基于 CRISPR 的通用免疫分析信号增强器 (CRUISE)。CRUISE 能够通过一抗或二抗集成到各种免疫测定中，灵敏度低至 1 fg mL^-1（~50 aM）和 6 个线性范围的对数，用于在 3-4 小时内检测细胞因子，例如 IFN-γ 和 EGFR。与商业 IFN-γ ELISA 试剂盒相比，它的灵敏度高出 10 ³倍，但使用相同的实验方案。对于人类血浆样品中的 IFN-γ 检测，实现了相同的 1 fg mL ^-1灵敏度以及 6 个对数的线性范围。我们期待我们的 CRUISE 为那些依赖免疫分析的人们提供一种替代但简单、用户友好和有效的策略，同时努力应对其灵敏度或检测范围的限制。