Since the emergence of coronavirus disease 2019 (COVID-19), this highly contagious disease has ravaged the world, cumulatively causing millions of deaths and huge economic losses. As the culprit of COVID-19, severe acute respiratory syndrome β-coronavirus 2 (SARS-CoV-2) is highly infectious and pathogenic, which has caused extreme panic worldwide. Early and rapid monitoring of SARS-CoV-2 has a significant role in suppressing the pandemic and reducing the virus's lethality. In our work, we prepared a self-enhanced ruthenium complex linked to zeolitic imidazole framework-8 (ZIF-8) and used it as an electrochemiluminescence (ECL) emitter. Additionally, a double-stranded specific nuclease (DSN)-assisted target RNA cycling with catalytic hairpin assembly (CHA) signal amplification technology was used to achieve the conversion of target RNA concentration to double-stranded DNA (dsDNA) output which significantly improved the detection sensitivity of target RNA under environmental conditions and in real human serum samples. In addition, we also combined the trans-cleavage property of CRISPR–Cas12a with the adsorption property of C₃N₄ on a ferrocene (Fc)-labeled DNA probe and obtained target RNA-dependent ECL signals. The reliable detection protocol achieved the transformation of SARS-CoV-2 RNA concentration to ECL responses, obtaining a limit of detection (LOD) of 0.67 fM with high specificity and reproducibility, which was of guiding significance for current detection methods of mutant SARS-CoV-2 and universal RNA.

中文翻译：

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CRISPR/Cas12a 介导的电化学发光平台用于环境和人血清 SARS-CoV-2 RNA 监测，使用与沸石咪唑框架 8 连接的自增强钌络合物

自2019冠状病毒病（COVID-19）出现以来，这种高度传染性疾病肆虐全球，累计造成数百万人死亡和巨大经济损失。作为COVID-19的罪魁祸首，严重急性呼吸综合征β-冠状病毒2（SARS-CoV-2）具有很强的传染性和致病性，在全世界引起了极度恐慌。对 SARS-CoV-2 的早期和快速监测对于抑制大流行和降低病毒的致死率具有重要作用。在我们的工作中，我们制备了一种与沸石咪唑骨架 8 (ZIF-8) 相连的自增强钌络合物，并将其用作电化学发光 (ECL) 发射器。此外，采用双链特异性核酸酶（DSN）辅助靶RNA循环和催化发夹组装（CHA）信号放大技术，实现靶RNA浓度向双链DNA（dsDNA）输出的转化，显着提高了检测灵敏度在环境条件下和真实的人血清样本中靶向 RNA。此外，我们还结合了CRISPR-Cas12a 的反式切割特性和 C ₃ N ₄对二茂铁 (Fc) 标记的 DNA 探针的吸附特性，并获得了目标 RNA 依赖性 ECL 信号。可靠的检测方案实现了 SARS-CoV-2 RNA 浓度向 ECL 反应的转化，获得了 0.67 fM 的检测限 (LOD)，具有高度的特异性和重现性，对目前的突变型 SARS-CoV 检测方法具有指导意义-2 和通用 RNA。