Nguyen Linh Viet, Giannetti Sara, Warren-Smith Stephen, Cooper Alan, Selleri Stefano, Cucinotta Annamaria, Monro Tanya
Institute for Photonics and Advanced Sensing (IPAS) and The ARC Centre for Nanoscale Biophotonics, The University of Adelaide, Adelaide 5005, Australia.
Information Engineering Department, University of Parma, Parma 43100, Italy.
Sensors (Basel). 2014 Aug 8;14(8):14488-99. doi: 10.3390/s140814488.
We report a novel approach to genotyping single nucleotide polymorphisms (SNPs) using molecular beacons in conjunction with a suspended core optical fiber (SCF). Target DNA sequences corresponding to the wild- or mutant-type have been accurately recognized by immobilizing two different molecular beacons on the core of a SCF. The two molecular beacons differ by one base in the loop-probe and utilize different fluorescent indicators. Single-color fluorescence enhancement was obtained when the immobilized SCFs were filled with a solution containing either wild-type or mutant-type sequence (homozygous sample), while filling the immobilized SCF with solution containing both wild- and mutant-type sequences resulted in dual-color fluorescence enhancement, indicating a heterozygous sample. The genotyping was realized amplification-free and with ultra low-volume for the required DNA solution (nano-liter). This is, to our knowledge, the first genotyping device based on the combination of optical fiber and molecular beacons.
我们报道了一种使用分子信标结合悬浮芯光纤(SCF)对单核苷酸多态性(SNP)进行基因分型的新方法。通过将两种不同的分子信标固定在SCF的芯上,已准确识别出对应野生型或突变型的目标DNA序列。这两种分子信标在环探针中相差一个碱基,并使用不同的荧光指示剂。当固定的SCF填充含有野生型或突变型序列的溶液(纯合样品)时,获得单色荧光增强,而用含有野生型和突变型序列的溶液填充固定的SCF则导致双色荧光增强,表明是杂合样品。该基因分型无需扩增,所需DNA溶液体积超低(纳升)。据我们所知,这是首个基于光纤和分子信标组合的基因分型装置。