A simple fluorescence aptasensor for gastric cancer exosome detection based on branched rolling circle amplification.
Rongrong HuangLei HeSong LiHongna LiuLian JinZhu ChenYongxiang ZhaoZhiyang LiYan DengNongyue HePublished in: Nanoscale (2020)
Exosomes are membrane nanovesicles carrying molecular information that may reflect the biological and genetic characteristics of their parent cells. Numerous studies have demonstrated the potential of exosomes as noninvasive cancer biomarkers. Hence, specific detection of cancer cell-derived exosomes is of significant importance. Here, we developed a fluorescence assay for the determination of gastric cancer exosomes based on branched rolling circle amplification (BRCA) and an aptamer to target specific exosomes. The designed padlock probe was cyclized after incubation with an aptamer binding with the target exosome. BRCA was triggered by adding a second primer and the resulting long tandem double-stranded DNA product was detected using SYBR Green I as the fluorescent dye. This method demonstrated a high specificity for target exosomes with a detection limit of 4.27 × 104 exosomes per mL. Moreover, plasma from gastric cancer patients was tested to verify the clinical applicability of this assay. Our results demonstrated that this aptamer-based biosensor may show potential for the early diagnosis of gastric cancer.
Keyphrases
- label free
- mesenchymal stem cells
- stem cells
- sensitive detection
- gold nanoparticles
- quantum dots
- single molecule
- loop mediated isothermal amplification
- papillary thyroid
- nucleic acid
- high throughput
- healthcare
- bone marrow
- gene expression
- living cells
- cell proliferation
- high resolution
- binding protein
- cell free
- circulating tumor
- childhood cancer
- highly efficient
- dna methylation