Obstetrics Department I, Northwest Women & Children's Hospital, Xi'an, Shaanxi Province, 710061, China.
Department of Obstetrics & Gynecology, Northwest Women & Children's Hospital, Xi'an, Shaanxi Province, 710061, China.
Biotechniques. 2023 Apr;74(4):172-178. doi: 10.2144/btn-2023-0008. Epub 2023 May 2.
miRNAs alter significantly throughout pregnancy to support the development of the fetus. However, sensitive detection of miRNA remains a challenge. Herein, a reliable miRNA detection approach integrating self-assembly-triggered signal amplification and CRISPR-Cas12a-system cleavage-based color generation is described. The colorimetric approach contains three signal amplification processes. The first signal amplification is formed by the released miRNA in a chain extension process. The produced sequence that is similar to the target miRNA initiates the second signal recycle. Finally, CRISPR-Cas12a-based transcleavage on linker sequences induces the third signal amplification. The method exhibits high sensitivity and a low limit of detection of 254 aM, showing promising prospects in disease diagnosis.
miRNAs 在整个怀孕期间会发生显著变化，以支持胎儿的发育。然而，miRNA 的灵敏检测仍然是一个挑战。在此，我们描述了一种可靠的 miRNA 检测方法，该方法集成了自组装触发的信号放大和基于 CRISPR-Cas12a 系统切割的颜色生成。该比色法包含三个信号放大过程。第一个信号放大是由释放的 miRNA 在链延伸过程中形成的。与靶 miRNA 相似的产生序列启动第二个信号循环。最后，基于 CRISPR-Cas12a 的连接序列的转切割诱导第三个信号放大。该方法表现出高灵敏度和低检测限为 254 aM，在疾病诊断中具有广阔的应用前景。
