State Key Lab of Electroanalytical Chemistry, Changchun Institute of Applied Chemistry, Chinese Academy of Sciences, Changchun, 130022, Jilin, China.
University of Science and Technology China, Hefei, 230026, Anhui, China.
Anal Bioanal Chem. 2023 May;415(12):2173-2183. doi: 10.1007/s00216-023-04637-3. Epub 2023 Mar 16.
Nucleic acid testing technology has made considerable progress in the last few years. However, there are still many challenges in the clinical application of multiple nucleic acid assays, such as how to ensure accurate results, increase speed and decrease cost. Herein, a three-way junction structure has been introduced to specifically translate analytes of loop-mediated isothermal amplification to a catalytic hairpin assembly. For different analyses, a well-optimized nucleic acid circuit can be directly applied to detection, through only one-component replacement, which only not avoids duplicate sequence design but also saves detection cost. Thanks to this design, multiple and logical analysis can be easily realized in a single reaction with ultra-high sensitivity and selectivity. In this paper, Mycoplasma pneumoniae and Streptococcus pneumoniae can be clearly distinguished from the clinical mixed sample with negative control or one analyte in one tube single fluorescence channel. The fair experimental results of actual clinical samples provide a strong support for the possibility of clinical application of this methodology.
核酸检测技术在过去几年中取得了相当大的进展。然而，在多重核酸检测的临床应用中仍然存在许多挑战，例如如何确保结果的准确性、提高速度和降低成本。在此，我们引入了一种三链结结构，将环介导等温扩增的分析物特异性地转化为催化发夹组装。对于不同的分析，只需通过单一组分替换，就可以将经过良好优化的核酸电路直接应用于检测，这不仅避免了重复的序列设计，还节省了检测成本。得益于这种设计，可以在单个反应中轻松实现多种逻辑分析，具有超高的灵敏度和选择性。在本文中，肺炎支原体和肺炎链球菌可以从临床混合样本中清晰区分，阴性对照或一管单荧光通道中的一个分析物即可实现。实际临床样本的公平实验结果为该方法的临床应用提供了有力支持。
