University of Parma, Department of Information Engineering, Area Parco delle Scienze 181/A, 43124, Parma, Italy.
J Biomed Opt. 2013 May;18(5):57004. doi: 10.1117/1.JBO.18.5.057004.
We describe a novel sensing approach based on a functionalized microstructured optical fiber-Bragg grating for specific DNA target sequences detection. The inner surface of a microstructured fiber, where a Bragg grating was previously inscribed, has been functionalized by covalent linking of a peptide nucleic acid probe targeting a DNA sequence bearing a single point mutation implicated in cystic fibrosis (CF) disease. A solution of an oligonucleotide (ON) corresponding to a tract of the CF gene containing the mutated DNA has been infiltrated inside the fiber capillaries and allowed to hybridize to the fiber surface according to the Watson-Crick pairing. In order to achieve signal amplification, ON-functionalized gold nanoparticles were then infiltrated and used in a sandwich-like assay. Experimental measurements show a clear shift of the reflected high order mode of a Bragg grating for a 100 nM DNA solution, and fluorescence measurements have confirmed the successful hybridization. Several experiments have been carried out on the same fiber using the identical concentration, showing the same modulation trend, suggesting the possibility of the reuse of the sensor. Measurements have also been made using a 100 nM mismatched DNA solution, containing a single nucleotide mutation and corresponding to the wild-type gene, and the results demonstrate the high selectivity of the sensor.
我们描述了一种基于功能化微结构光纤布拉格光栅的新型传感方法,用于特定 DNA 靶序列的检测。在先前刻写布拉格光栅的微结构光纤的内表面上,通过共价连接针对单核苷酸点突变的肽核酸探针来进行功能化,该突变与囊性纤维化 (CF) 疾病相关。与包含突变 DNA 的 CF 基因片段相对应的寡核苷酸 (ON) 的溶液已渗透到光纤毛细管内部,并根据沃森-克里克配对原则与纤维表面杂交。为了实现信号放大,然后渗透了 ON 功能化的金纳米粒子,并在夹心样测定中使用。实验测量显示,对于 100 nM 的 DNA 溶液,布拉格光栅的反射高阶模式发生了明显的偏移,并且荧光测量已经证实了成功的杂交。在同一根光纤上使用相同的浓度进行了几次实验,显示出相同的调制趋势,这表明传感器可以重复使用。使用包含单个核苷酸突变且对应于野生型基因的 100 nM 错配 DNA 溶液进行了测量,结果表明传感器具有高选择性。