特异性非标记适体光谱椭偏法检测前列腺癌生物标志物PCA3:与电化学检测的比较

S. Takita, A. Nabok, David P. Smith, A. Lishchuk
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引用次数: 1

摘要

最常见的前列腺癌(PCa)诊断是基于血液中前列腺特异性抗原(PSA)的检测,具有特异性限制,通常导致假阳性和假阴性结果;因此,使用更具体的前列腺癌生物标志物来改善前列腺癌诊断是非常重要的。研究表明,前列腺癌患者尿液中过表达的长链非编码RNA前列腺癌抗原3 (lncPCA3)是前列腺癌无创早期诊断的理想生物标志物。基于适体生物受体(aptassensors)的基因传感器为检测PCa生物标志物提供了成本和时间效益高、精确的诊断工具。在这项研究中,我们报告了利用光学(光谱椭偏)测量与早期发表的电化学(CV和IS)测量相比较的基于rna的适体传感器的进一步发展。这些传感器是通过将巯基化CG-3 RNA适配体固定在金表面制成的。代替之前电化学测量中使用的氧化还原标记适配体,这里使用了非标记适配体与全内反射椭偏仪(TIRE)测量相结合。比较了两种方法的结果。TIRE方法具有潜在的高度敏感性,在这方面可与能够检测亚pm浓度水平的PCA3的电化学方法相媲美。所需的选择性是由pca3 -适体与KD结合在10−9 M范围内的高亲和力提供的。光谱椭偏测量提供了PCA3与适体结合过程的额外信息。
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Spectroscopic Ellipsometry Detection of Prostate Cancer Bio-Marker PCA3 Using Specific Non-Labeled Aptamer: Comparison with Electrochemical Detection
The most common prostate cancer (PCa) diagnostics, which are based on detection of prostate-specific antigens (PSA) in blood, have specificity limitations often resulting in both false-positive and false-negative results; therefore, improvement in PCa diagnostics using more specific PCa biomarkers is of high importance. Studies have shown that the long noncoding RNA Prostate Cancer Antigen 3 (lncPCA3) that is over-expressed in the urine of prostate cancer patients is an ideal biomarker for non-invasive early diagnostics of PCa. Geno-sensors based on aptamer bioreceptors (aptasensors) offer cost- and time-effective, and precise diagnostic tools for detecting PCa biomarkers. In this study, we report on further developments of RNA-based aptasensors exploiting optical (spectroscopic ellipsometry) measurements in comparison with electrochemical (CV and IS) measurements published earlier. These sensors were made by immobilization of thiolated CG-3 RNA aptamers on the surface of gold. Instead of a redox-labelled aptamer used previously in electrochemical measurements, a non-labelled aptamer was used here in a combination with total internal reflection ellipsometry (TIRE) measurements. The results obtained by these two methods were compared. The method of TIRE is potentially highly sensitive and comparable in that respect with electrochemical methods capable of detection of PCA3 in sub-pM levels of concentration. The required selectivity is provided by the high affinity of PCA3-to-aptamer binding with KD in the 10−9 M range. The spectroscopic ellipsometry measurements provided additional information on the processes of PCA3 to aptamer binding.
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