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癌细胞中核酸和端粒酶活性的磁机械检测

Magneto-mechanical detection of nucleic acids and telomerase activity in cancer cells.

作者信息

Weizmann Yossi, Patolsky Fernando, Lioubashevski Oleg, Willner Itamar

机构信息

Institute of Chemistry, The Hebrew University of Jerusalem, Jerusalem 91904, Israel.

出版信息

J Am Chem Soc. 2004 Feb 4;126(4):1073-80. doi: 10.1021/ja038257v.

Abstract

The ultra-sensitive magneto-mechanical detection of DNA, single-base-mismatches in nucleic acids, and the assay of telomerase activity are accomplished by monitoring the magnetically induced deflection of a cantilever functionalized with magnetic beads associated with the biosensing interface. The analyzed M13phi DNA hybridized with the nucleic acid-functionalized magnetic beads is replicated in the presence of dNTPs that include biotin-labeled dUTP. The resulting beads are attached to an avidin-coated cantilever, and the modified cantilever is deflected by an external magnetic field. Similarly, telomerization of nucleic acid-modified magnetic beads in the presence of dNTPs, biotin-labeled dUTP, and telomerase from cancer cell extracts and the subsequent association of the magnetic beads to the cantilever surface results in the lever deflection by an external magnetic field. M13phi DNA is sensed with a sensitivity limit of 7.1 x 10(-20) M by the magneto-mechanical detection method.

摘要

通过监测与生物传感界面相关联的磁珠功能化悬臂的磁致偏转,可实现对DNA、核酸中的单碱基错配以及端粒酶活性的超灵敏磁机械检测。与核酸功能化磁珠杂交的分析型M13phi DNA在包含生物素标记的dUTP的dNTP存在下进行复制。所得磁珠附着于抗生物素蛋白包被的悬臂上,经修饰的悬臂会因外部磁场而发生偏转。同样,在dNTP、生物素标记的dUTP以及癌细胞提取物中的端粒酶存在的情况下,核酸修饰磁珠的端粒化以及随后磁珠与悬臂表面的结合会导致杠杆在外部磁场作用下发生偏转。采用磁机械检测方法检测M13phi DNA的灵敏度极限为7.1×10(-20) M。

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