The ultrasensitive sensor with three optical response mechanisms was proposed for the determination of trace amounts of norovirus using a 3-in-1 GCSNAs (a gap-containing spherical nucleic acid nanoparticles) probe. A simple and highly sensitive three-mode biosensor with Raman, colorimetric, and fluorescence functions was proposed and implemented using the GCSNAs probe and a DNA hydrogel for norovirus detection. When the virus exists, the trans-cleavage activity of CRISPR-Cas12a was activated by double-stranded dsDNA (dsDNA) generated by reverse transcription and recombinase polymerase isothermal amplification (RT-RPA) to degrade the DNA hydrogel/GCSNA composition and release the three-in-one (3-in-1) probe-GCSNA, realising the triple ultrasensitive detection of norovirus. The colorimetric sensing mode allows for semi-quantitative on-site detection, which is visible to the naked eye and the quantitative detection can be achieved by conducting grayscale analysis using the "Colour Grab" function of a smartphone. This new triple sensor achieved the successful quantification of norovirus at concentrations as low as the femtomolar scale with an excellent selectivity and accuracy. Considering the colorimetric properties of rolling circle amplification (RCA)-based DNA hydrogels and GCSNAs, the proposed method has a broad application prospect in virus on-site detection in food. It should be applicable for virus detection in a wide range of fields such, as environmental analysis, medical diagnosis, and food safety. It is anticipated that this mechanism will open new avenues for the development of multimodal analyses and multifunctional sensing platforms for various applications. We anticipate that this sensing mechanism will open up a new way for the development of food safety detection.

中文翻译：

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基于DNA水凝胶和复合功能纳米材料的三合一CRISPR平台对诺如病毒进行超灵敏检测


提出了具有三种光学响应机制的超灵敏传感器，用于使用 3 合 1 GCSNAs（一种包含间隙的球形核酸纳米颗粒）探针测定痕量诺如病毒。提出了一种简单且高灵敏度的具有拉曼、比色和荧光功能的三模式生物传感器，并使用 GCSNAs 探针和 DNA 水凝胶实现诺如病毒检测。当病毒存在时，CRISPR-Cas12a 的反式切割活性被逆转录和重组酶聚合酶等温扩增 （RT-RPA） 产生的双链 dsDNA （dsDNA） 激活，降解 DNA 水凝胶/GCSNA 组成并释放三合一 （3-in-1） 探针-GCSNA，实现诺如病毒的三重超灵敏检测。比色传感模式允许进行肉眼可见的半定量现场检测，并且可以通过使用智能手机的“颜色抓取”功能进行灰度分析来实现定量检测。这种新型三重传感器成功定量了低至飞摩尔级浓度的诺如病毒，具有出色的选择性和准确性。考虑到基于滚环扩增 （RCA） 的 DNA 水凝胶和 GCSNAs 的比色特性，所提方法在食品中病毒现场检测中具有广阔的应用前景。它应该适用于环境分析、医学诊断和食品安全等广泛领域的病毒检测。预计该机制将为开发用于各种应用的多模态分析和多功能传感平台开辟新途径。 我们预计这种传感机制将为食品安全检测的发展开辟一条新的途径。