基于DNA的扩增电学生物条形码分析法用于一锅法检测两种靶标DNA。
DNA-based amplified electrical bio-barcode assay for one-pot detection of two target DNAs.
作者信息
Zhang Xiaoru, Su Haoran, Bi Sai, Li Shuguo, Zhang Shusheng
机构信息
Key Laboratory of Eco-chemical Engineering, Ministry of Education, College of Chemistry and Molecular Engineering, Qingdao University of Science and Technology, Qingdao 266042, PR China.
出版信息
Biosens Bioelectron. 2009 Apr 15;24(8):2730-4. doi: 10.1016/j.bios.2008.12.032. Epub 2008 Dec 30.
A sensitive label-free bio-barcode assay provided a PCR-free method for quantitative detection of two nucleic acid targets (HTLV-I and HTLV-II) simultaneously. This DNA biosensor was fabricated with two-component oligonucleotide-modified gold nanoparticles (AuNPs) and two-component oligonucleotide-modified magnetic beads (MBs), which can sandwich a specific target. After liberating the adsorbed thiolated barcode DNA strands (poly A and poly G) from the AuNPs surface with dithiothreitol (DTT) and acidic dipurinization, the electrochemical measurements were directly performed based on the redox activity of guanine (G) and adenine (A) nucleobases. Under the optimal assembling and detection conditions, a good linearity for simultaneous detection was obtained in the range from 4.4x10(-11) to 2.0x10(-9) M, and the detection limit (3sigma) was estimated to be 1.71x10(-12) M for T(1)-DNA and 1.55x10(-12) M for T(2)-DNA.
一种灵敏的无标记生物条形码检测法提供了一种无需聚合酶链反应（PCR）的方法，可同时定量检测两种核酸靶标（人类嗜T淋巴细胞病毒I型[HTLV-I]和人类嗜T淋巴细胞病毒II型[HTLV-II]）。这种DNA生物传感器由两组分寡核苷酸修饰的金纳米颗粒（AuNPs）和两组分寡核苷酸修饰的磁珠（MBs）构建而成，能够夹合特定靶标。在用二硫苏糖醇（DTT）和酸性脱嘌呤作用从AuNPs表面释放吸附的硫醇化条形码DNA链（聚A和聚G）后，基于鸟嘌呤（G）和腺嘌呤（A）核碱基的氧化还原活性直接进行电化学测量。在最佳组装和检测条件下，同时检测的线性良好，范围为4.4×10⁻¹¹至2.0×10⁻⁹ M，T(1)-DNA的检测限（3σ）估计为1.71×10⁻¹² M，T(2)-DNA的检测限为1.55×10⁻¹² M。
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