Functional Molecules Analysis and Biotransformation Key Laboratory of Universities in Yunnan Province, School of Chemical Science and Technology, Yunnan University, Kunming, China.
Hunan Province Geological Testing Institute, Changsha, China.
Luminescence. 2020 Aug;35(5):702-708. doi: 10.1002/bio.3775. Epub 2020 Jan 11.
Recognition and quantification of oligonucleotide sequences play important roles in medical diagnosis. In this study, a new fluorescent oligonucleotide-stabilized silver nanocluster beacon (NCB) probe was designed for sensitive detection of oligonucleotide sequence targets. This probe contained two tailored DNA strands. One strand was a signal probe strand containing a cytosine-rich strand template for fluorescent silver nanocluster (Ag NC) synthesis and a detection sections at each end. The other strand was a fluorescence enhancing strand containing a guanine-rich section for signal enhancement at one end and a linker section complementary to one end of the signal probe strand. After synthesis of the Ag NCs and hybridization of the two strands, the fluorescence intensity of the as-prepared silver NCB was enhanced 200-fold compared with the Ag NCs. Two NCBs were designed to detect two disease-related oligonucleotide sequences, and results indicated that the two target oligonucleotide sequences in the range 50.0-600.0 and 50.0-200.0 nM could be linearly detected with detection limits of 20 and 25 nM, respectively. The developed fluorescence method using NCBs for oligonucleotide sequence detection was sensitive, facile and had potential for use in bioanalysis and diagnosis.
寡核苷酸序列的识别和定量在医学诊断中具有重要作用。在这项研究中,设计了一种新的荧光寡核苷酸稳定的银纳米簇荧光探针(NCB),用于灵敏检测寡核苷酸序列靶标。该探针包含两条定制的 DNA 链。一条链是信号探针链,包含一个富含胞嘧啶的链模板,用于荧光银纳米簇(Ag NC)的合成,两端各有一个检测部分。另一条链是荧光增强链,在一端含有富含鸟嘌呤的部分,用于信号增强,在另一端有一个与信号探针链一端互补的连接部分。合成 Ag NC 并杂交两条链后,与 Ag NC 相比,制备的银 NCB 的荧光强度增强了 200 倍。设计了两个 NCB 来检测两个与疾病相关的寡核苷酸序列,结果表明,两个靶标寡核苷酸序列在 50.0-600.0 和 50.0-200.0 nM 范围内可以线性检测,检测限分别为 20 和 25 nM。该研究开发的使用 NCB 进行寡核苷酸序列检测的荧光方法具有灵敏度高、简便的特点,具有用于生物分析和诊断的潜力。