Magneto-mechanical detection of nucleic acids and telomerase activity in cancer cells
Magneto-mechanical detection of nucleic acids and telomerase activity in cancer cells
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DOI：
10.1021/ja038257v
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发表时间：
2004-02-04
影响因子：
15
通讯作者：
Willner, I
中科院分区：
文献类型：
--
作者：
Weizmann, Y;Patolsky, F;Willner, I
DNA的超灵敏磁力检测、核酸中的单碱基错配和端粒酶活性的测定是通过监测与生物传感界面相关的磁珠功能化的悬臂的磁感应偏转来完成的。分析的M13phi DNA与核酸功能化磁珠杂交，在dNTPs存在下复制，其中dNTPs包括生物素标记的dUTP。得到的珠子被固定在一个涂有亲和素的悬臂梁上，修改后的悬臂梁在外部磁场的作用下发生偏转。同样，核酸修饰的磁珠在dNTPs、生物素标记的dUTP和癌细胞提取物中的端粒酶存在下的端粒化以及随后的磁珠与悬臂表面的结合导致外部磁场导致杠杆偏转。磁机检测M13phi DNA的灵敏度极限为7.1×10(-20)M。
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.