Ultrasensitive detection of miRNA with single-nucleotide resolution using a high-electron-mobility transistor biosensor combined with splint-ligation
Chenyang Yang (Chinese Academy of Sciences)
Jingya Tang (Chinese Academy of Sciences)
Jianwen Sun (Tsinghua University)
Zewen Liu (Tsinghua University)
Guoqi Zhang (TU Delft - Electronic Components, Technology and Materials)
Zhe Li (Precision Scientific (Beijing) Co. Ltd.)
Dian Bing Wang (Chinese Academy of Sciences)
Xian En Zhang (Student TU Delft, Shenzhen University of Advanced Technology, Chinese Academy of Sciences)
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Abstract
The use of microRNAs as clinical cancer biomarkers is hindered by the absence of accurate, sensitive and rapid assays for their detection in biofluids. Here we report a biosensing approach, SpLig-HEMT, that combines an RNA splint-ligation reaction with an AlGaN/GaN high-electron-mobility transistor biosensor for ultrasensitive miRNA detection. In this system, HEMT functions as a highly effective voltage amplifier to enhance detection sensitivity, while the splint-ligation reaction ensures precise discrimination of single-nucleotide mutations. By detecting miRNA-21, the SpLig-HEMT biosensor achieves an exceptional limit of detection of 10−18 M within 30 min, with a dynamic range from 10−18 M to 10−13 M. No detectable response is observed for one-mismatch miR-21. Furthermore, the SpLig-HEMT biosensor enables direct analysis of blood serum samples, effectively distinguishing between healthy individuals and patients with ovarian cancer. This study addresses critical challenges in miRNA detection and presents a promising tool for cancer diagnosis and prognosis.
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