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Fluorescent aptasensor based on G-quadruplex-assisted structural transformation for the detection of biomarker lipocalin 1.
Biosensors and Bioelectronics ( IF 12.6 ) Pub Date : 2020-09-10 , DOI: 10.1016/j.bios.2020.112607
Shunxiang Gao 1 , Shenghai Zhang 1 , Xinghuai Sun 1 , Xin Zheng 2 , Jihong Wu 1
Affiliation  

Diabetic retinopathy (DR) is the leading global cause of blindness in the working-age population. Early diagnosis and intervention can effectively reduce the risk for blindness. However, the current diagnostic methods in clinical practice remain constrained by nonquantitative examinations and individual ophthalmologists’ experiences. Sensitive, specific and accurate detection of DR-specific biomarkers is an important approach to achieve its early and rapid diagnosis. In this study, a high-affinity aptamer APT12TM that specifically binds to the tear-derived DR biomarker lipocalin 1 was obtained. The aptamer APT12TM can be folded into a stable B-DNA structure, and its strong interaction with LCN 1, including hydrogen bonding and hydrophobic interactions, is an important factor for targeted recognition and high-affinity binding. A G-rich DNA fragment was further assembled at both ends of the aptamer APT12TM, and the B-DNA form was successfully converted into a parallel G-quadruplex. Most importantly, LCN 1 could induce further transformation of the G-quadruplex structure. Therefore, a fluorescent aptasensor based on G-quadruplex-assisted structural transformation was developed through the Thioflavin T mediator. The aptasensor exhibited a broad detection window from 0.25 to 1000 nM LCN 1, with a limit of detection of 0.2 nM. Furthermore, the aptasensor was applied to LCN 1 detection in artificial tear samples and displayed good reproducibility and stability. These results show that the developed aptasensor has significant potential for sensitive, specific and convenient detection of the DR-specific biomarker LCN 1.



中文翻译:

基于G-四链体辅助结构转化的荧光适体传感器,用于检测生物标志物lipocalin 1。

糖尿病性视网膜病(DR)是全球工作年龄段失明的主要原因。早期诊断和干预可以有效减少失明的风险。但是,当前临床实践中的诊断方法仍然受到非定量检查和个别眼科医生经验的限制。灵敏,特异和准确地检测DR特异性生物标志物是实现其早期和快速诊断的重要方法。在这项研究中,获得了一种高亲和力的适体APT12TM,它与泪液衍生的DR生物标志物lipocalin 1特异性结合。适体APT12TM可以折叠成稳定的B-DNA结构,它与LCN 1的强相互作用,包括氢键和疏水相互作用,是靶向识别和高亲和力结合的重要因素。在适体APT12TM的两端进一步组装了一个富含G的DNA片段,并将B-DNA形式成功转化为平行的G-四链体。最重要的是,LCN 1可以诱导G-四链体结构的进一步转化。因此,通过硫黄素T介体开发了基于G-四链体辅助的结构转变的荧光适体传感器。适体传感器显示了从0.25到1000 nM LCN 1的宽检测窗,检测限为0.2 nM。此外,将适体传感器应用于人工泪液样品中的LCN 1检测,并显示出良好的重现性和稳定性。这些结果表明,开发的适体传感器具有灵敏,特异和方便地检测DR特异性生物标志物LCN 1的巨大潜力。B-DNA形式成功转化为平行的G-四链体。最重要的是,LCN 1可以诱导G-四链体结构的进一步转化。因此,通过硫黄素T介体开发了基于G-四链体辅助的结构转变的荧光适体传感器。适体传感器显示了从0.25到1000 nM LCN 1的宽检测窗,检测限为0.2 nM。此外,将适体传感器应用于人工泪液样品中的LCN 1检测,并显示出良好的重现性和稳定性。这些结果表明,开发的适体传感器具有灵敏,特异和方便地检测DR特异性生物标志物LCN 1的巨大潜力。B-DNA形式成功转化为平行的G-四链体。最重要的是,LCN 1可以诱导G-四链体结构的进一步转化。因此,通过硫黄素T介体开发了基于G-四链体辅助的结构转变的荧光适体传感器。适体传感器显示了从0.25到1000 nM LCN 1的宽检测窗,检测限为0.2 nM。此外,将适体传感器应用于人工泪液样品中的LCN 1检测,并显示出良好的重现性和稳定性。这些结果表明,开发的适体传感器具有灵敏,特异和方便的DR特异性生物标志物LCN 1检测潜力。LCN 1可以诱导G四联体结构的进一步转化。因此,通过硫黄素T介体开发了基于G-四链体辅助的结构转变的荧光适体传感器。适体传感器显示了从0.25到1000 nM LCN 1的宽检测窗,检测限为0.2 nM。此外,将适体传感器应用于人工泪液样品中的LCN 1检测,并显示出良好的重现性和稳定性。这些结果表明,开发的适体传感器具有灵敏,特异和方便的DR特异性生物标志物LCN 1检测潜力。LCN 1可以诱导G四联体结构的进一步转化。因此,通过硫黄素T介体开发了基于G-四链体辅助的结构转变的荧光适体传感器。适体传感器显示了从0.25到1000 nM LCN 1的宽检测窗,检测限为0.2 nM。此外,将适体传感器应用于人工泪液样品中的LCN 1检测,并显示出良好的重现性和稳定性。这些结果表明,开发的适体传感器具有灵敏,特异和方便的DR特异性生物标志物LCN 1检测潜力。25至1000 nM LCN 1,检出限为0.2 nM。此外,将适体传感器应用于人工泪液样品中的LCN 1检测,并显示出良好的重现性和稳定性。这些结果表明,开发的适体传感器具有灵敏,特异和方便的DR特异性生物标志物LCN 1检测潜力。25至1000 nM LCN 1,检出限为0.2 nM。此外,将适体传感器应用于人工泪液样品中的LCN 1检测,并显示出良好的重现性和稳定性。这些结果表明,开发的适体传感器具有灵敏,特异和方便的DR特异性生物标志物LCN 1检测潜力。

更新日期:2020-09-15
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