Ultrasensitive Biosensors Detecting m6A in Blood: Achieving Early Screening and Typing of Tumors

血型 打字 生物传感器 化学 医学 纳米技术 生物 材料科学 遗传学 免疫学
作者
Haiping Li,Xixiang Xie,Xiyu Liu,Pan Wu,Jian He,Faquan Lin,Liang Shi,Yong Huang
出处
期刊:ACS Sensors [American Chemical Society]
标识
DOI:10.1021/acssensors.4c01875
摘要

N6-methyladenosine (m6A) modification is one of the most widespread RNA modifications in eukaryotes and is involved in cancer development and progression by regulating oncogene expression. Herein, a reticulated rolling circle amplification (RCA) cascade reaction was used to construct a novel electrochemical biosensor for ultrasensitive detection of m6A, employing ferrocene-tyramine (Fc-Tyr) molecules as electroactive probes. In this strategy, the RCA cascade reaction not only amplifies specific circular DNA in the designed template to reduce the binding with similar nucleic acid sequences but also generates a long ssDNA through multiple repetitions to capture a large number of electrochemical signal probes and achieve the amplification of electrochemical biosensing signals. The developed biosensor demonstrated high selectivity and sensitivity toward m6A in the range of 0.5 pM-150 nM, with a detection limit of 14.07 fM. Meanwhile, total RNA extracted from cell samples was analyzed for m6A expression levels using the developed biosensor and a commercial colorimetric immunoassay, the biosensor and immunoassay showed consistent results. In addition, m6A levels in clinical serum samples were assessed using the developed electrochemical biosensor, which showed that m6A expression was much lower in healthy individuals than in cancer patients, therefore the biosensor is promising for cancer typing. This study provides a new method for rapid and convenient tumor marker detection in clinical practice, as well as a new idea for sensitive detection of other biomolecules.
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