Beijing Advanced Innovation Center for Food Nutrition and Human Health, Beijing Technology and Business University (BTBU), Beijing, 102488, China; School of Life Sciences, Central South University, Changsha, 410013, China.
School of Life Sciences, Central South University, Changsha, 410013, China.
Mol Cell Probes. 2019 Aug;46:101423. doi: 10.1016/j.mcp.2019.101423. Epub 2019 Jul 16.
Polydeoxyadenosine (poly (dA)) has been extensively applied for detecting many drug molecules. Herein, we developed a sensitive method for detecting coralyne and heparin using a modified DNA probe with poly (dA) at one end. In the absence of coralyne, the DNA probe was digested by the Exonuclease I (Exo I), and therefore the SYBR Green I (SG I) emitted an extremely low fluorescent signal. While coralyne specifically binding to poly (dA) with strong propensity could remarkably restrain the disintegration of the DNA probe, through which as a template the second strand of DNA sequence was formed with the introduction of DNA polymerase. Therefore, the fluorescent signal of SG I was intensified to quantify coralyne. Based on this method, heparin can be determined due to its strong affinity towards coralyne. This method showed a linear range from 2 to 500 nM for coralyne with a low detection limit of 0.98 nM, and the linear range of heparin was from 1 to 100 nM when 1.25 nm was the detection limit. The proposed method was also implemented successfully in biological samples and showed a potential application for screening potential therapeutic molecules.
聚脱氧腺苷（poly(dA)）已广泛应用于检测多种药物分子。在此，我们开发了一种使用一端带有聚（dA）的修饰 DNA 探针检测珊瑚萘和肝素的灵敏方法。在没有珊瑚萘的情况下，DNA 探针被内切酶 I（Exo I）消化，因此 SYBR Green I（SG I）发出极低的荧光信号。而珊瑚萘特异性地与聚（dA）结合，具有很强的倾向性，可以显著抑制 DNA 探针的解体，通过该探针作为模板，引入 DNA 聚合酶形成第二条 DNA 序列。因此，SG I 的荧光信号增强以定量珊瑚萘。基于这种方法，由于肝素对珊瑚萘具有很强的亲和力，因此可以对其进行测定。该方法对珊瑚萘的线性范围为 2 至 500 nM，检测限低至 0.98 nM，当检测限为 1.25 nM 时，肝素的线性范围为 1 至 100 nM。该方法还成功地应用于生物样品中，显示出用于筛选潜在治疗分子的应用潜力。
