DNA hybridization detection with 100 zM sensitivity using piezoelectric plate sensors with an improved noise-reduction algorithm.
DNA hybridization detection with 100 zM sensitivity using piezoelectric plate sensors with an improved noise-reduction algorithm.
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DOI：
10.1039/c4an00215f
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发表时间：
2014-06-07
期刊：
影响因子：
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通讯作者：
Shih WY
中科院分区：
文献类型：
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作者：
Kirimli CE;Shih WH;Shih WY
我们研究了实时原位杂交检测缓冲溶液和尿液中的靶DNA（tDNA），使用8 μ m厚的PMN-PT压电板传感器（piezoelectric plate sensors，PEPS），长约1.1-1.2 mm，宽约0.45 mm，改进的3-巯基丙基三甲氧基硅烷（MPS）绝缘和一种新的多抛物线（>50）共振峰位置拟合算法。利用固定在PEPS表面上的探针DNA（pDNA）并通过监测第一宽度延伸模式（韦姆）共振频率偏移，我们在真实的时间内检测尿液（100 zM）中浓度低至1 × 10−19 M的tDNA，信噪比（SNR）为13，而无需在室温下30分钟内进行DNA分离和扩增。抛物线拟合算法将SNR的检测与使用原始数据获得的SNR的检测相比增加了约10倍，并且与使用单抛物线拟合获得的SNR的检测相比增加了约5倍。通过原位跟踪检测和随后的荧光报告微球（FRM）的可视化进行验证，所述荧光报告微球涂覆有与tDNA互补但不同于探针pDNA的报告DNA。
We have examined real-time, in situ hybridization detection of target DNA (tDNA) in a buffer solution and in urine using 8 μm-thick lead magnesium niobate–lead titanate (PMN–PT) piezoelectric plate sensors (PEPSs) about 1.1–1.2 mm long and 0.45 mm wide with improved 3-mercaptopropyltrimethoxysilane (MPS) insulation and a new multiple-parabola (>50) resonance peak position fitting algorithm. With probe DNA (pDNA) immobilized on the PEPS surface and by monitoring the first width extension mode (WEM) resonance frequency shift we detected tDNA in real time at concentration as low as 1 × 10−19 M in urine (100 zM) with a signal to noise ratio (SNR) of 13 without DNA isolation and amplification at room temperature in 30 min. The present multiple-parabola fitting algorithm increased the detection of SNR by about 10 times compared to those obtained using the raw data and by about 5 times compared to those obtained using single parabola fitting. The detection was validated by in situ follow-up detection and subsequent visualization of fluorescent reporter microspheres (FRMs) coated with reporter DNA complementary to the tDNA but different from the probe pDNA.