MicroRNAs are proposed novel biomarker for noninvasive diagnosis of cancer. miRNA-143 is reported to be associated with the development of prostate cancer. However, detection of miRNAs is still challenging due to their unique characteristics, such as small size and high sequence homology among family members. We here developed a gold nanoparticle (AuNP)-based visual assay that combines with CRISPR/Cas12a-assisted hyperbranched rolling circle amplification (HRCA), which is called HRCA enhanced CRISPR/Cas12a-based assay (HECA) for sensitive detection of miRNA-143. The sequence-specific recognition character of CRISPR/Cas12a and HRCA signal amplification strategy enables the HECA outstanding specificity and sensitivity. In optimal condition, 1 fM miRNA-143 could be detected by naked eyes, and down to aM level with the aid of UV–Vis instrument. The diagnostic performance of the HECA for clinical samples was also evaluated based on the receiving operating characteristic algorithm (ROC), and our results suggest the miR-143 is a promising biomarker for noninvasive diagnosis of prostate cancer. This method is simple in operation and requires minimum instrument. We expect it to be widely applied in clinical diagnostics, especially in low-resource settings.

微小RNA被提出用于癌症的非侵入性诊断的新型生物标志物。据报道，miRNA-143 与前列腺癌的发展有关。然而，由于 miRNA 具有独特的特征，例如体积小、家族成员之间的序列同源性高，检测 miRNA 仍然具有挑战性。我们在这里开发了一种基于金纳米颗粒 (AuNP) 的视觉检测方法，该检测方法与 CRISPR/Cas12a 辅助的超支化滚环扩增 (HRCA) 相结合，称为 HRCA 增强的基于 CRISPR/Cas12a 的检测方法 (HECA)，用于灵敏检测 miRNA-143 . CRISPR/Cas12a的序列特异性识别特性和HRCA信号放大策略使HECA具有出色的特异性和灵敏度。在最佳条件下，肉眼可检测到1 fM miRNA-143，借助UV-Vis仪器可检测到aM水平。HECA 对临床样本的诊断性能也基于接收操作特征算法 (ROC) 进行了评估，我们的结果表明 miR-143 是前列腺癌无创诊断的有前途的生物标志物。该方法操作简单，所需仪器最少。我们预计它将广泛应用于临床诊断，尤其是在资源匮乏的环境中。