Label-free detection of HPV mRNA with an artificial chaperone-enhanced MNAzyme (ACEzyme)-based electrochemical sensor
Label-free detection of HPV mRNA with an artificial chaperone-enhanced MNAzyme (ACEzyme)-based electrochemical sensor
复制标题
使用基于人工伴侣增强 MNAzyme (ACEzyme) 的电化学传感器对 HPV mRNA 进行无标记检测
DOI：
10.1016/j.bios.2022.114352
复制
发表时间：
2023
影响因子：
12.6
通讯作者：
Ngamrojanavanich Nattaya
中科院分区：
文献类型：
--
作者：
Hanpanich Orakan;Lomae Atchara;Maruyama Atsushi;Palaga Tanapat;Chailapakul Orawon;Ngamrojanavanich Nattaya
用于医疗(POC)诊断应用的核酸生物传感器是非常理想的。能够以简单、快速、负担得起和便携的形式检测DNA和RNA，导致在疾病监测和管理领域的早期筛查的一系列重要应用。在此，我们报道了一种基于靶向驱动的MNAzyme切割活性的恒温、无标记的电化学生物传感器的开发。与HPV mRNA的杂交，一个模式的核酸靶标，激活MNAzyme并启动固定发夹底物的切割，导致电化学信号的变化。在最佳条件下，该方法的检出限为2.6 pm，孵化时间为60min。此外，首次将人工伴侣增强型酶(ACEzyme)系统集成到电化学生物传感器中。PLL-g-Dex的加入提高了生物传感器的分析性能，显著缩短了检测时间，具有核酸伴侣活性。在没有预先扩增的情况下，孵育时间缩短了三倍，检测下限为0.88 pm。提出的生物传感平台具有制造和操作简单、在存在单碱基错配的情况下具有良好的选择性以及在复杂的总RNA混合物中具有良好的通用性等优点。我们相信，这个恒温、无标记、无蛋白质的核酸分析平台可以为进一步开发作为临床应用的通用核酸生物传感平台提供基础。
Nucleic acid biosensors for point-of-care (POC) diagnostic applications are highly desirable. The ability to detect DNA and RNA in a simple, rapid, affordable and portable format leads to a range of important applications for early screening in the field of disease monitoring and management. Herein, we report the development of an isothermal, label-free electrochemical biosensor that was designed on the basis of target-driven MNAzyme cleavage activity. Hybridization with HPV mRNA, a model nucleic acid target, activated MNAzyme and initiated the cleavage of immobilized hairpin substrates, leading to changes in the electrochemical signal. Under optimal conditions, a detection limit of 2.6 pM was obtained with an incubation time of 60 min. Furthermore, an artificial chaperone-enhanced MNAzyme (ACEzyme) system was integrated to an electrochemical biosensor for the first time. The analytical performance of the biosensor was enhanced, and the detection time was significantly reduced by the addition of PLL-g-Dex, which exhibits nucleic acid chaperone-like activity. A detection limit of 0.88 pM was obtained with a threefold decrease in incubation time without prior amplification. The proposed biosensing platform shows the advantages of simple fabrication and operation, good selectivity in the presence of single-base mismatch, and excellent versatility in a complex mixture of total RNA. We believe that this isothermal, label-free, and protein-free nucleic acid analysis platform could provide foundations for the further development of a universal nucleic acid biosensing platform for clinical application.