MicroRNA Detection with Turnover Amplification via Hybridization-Mediated Staudinger Reduction for Pancreatic Cancer Diagnosis.
Liman XianFeng XuJianzhou LiuNing XuHaidong LiHaoying GeKun ShaoJiangli FanGuishan XiaoXiaojun PengPublished in: Journal of the American Chemical Society (2019)
The occurrence of and development in the early pathological stage of pancreatic cancer has proved to be associated with microRNAs. However, it remains a great challenge to directly monitor low-expression, and downregulation of, microRNA among living cells, tissues, and serum samples. In this work, Staudinger reduction is first applied in intracellular microRNA detection, establishing a set of smart hybridization-mediated Staudinger reduction probes (HMSR-probe) which contain designed oligonucleotide sequences. Meanwhile, 40 serum samples (healthy people (6), patients with pancreatitis (22), and pancreatic cancer patients (12)) are tested for exploring the potential clinical application. Of note, the molecules bound to nucleic acid confine the reactive site to close proximity in a compact space, and nonconnected product from Staudinger reaction facilitates turnover amplification to an ameliorative detection limit (1.3 × 10-15 M). Moreover, compared with qRT-PCR, a low false positive signal and an excellent specificity makes the probe more suitable and convenient for pancreatic cancer diagnosis in blood samples. For practical applications, HMSR-probe enable accurate differentiation in cell and tissue samples under both 488 and 785 nm and have good coherence to known research. As a proof of concept, the reliable results in distinguishing pancreatic cancer patients from different morbid stages might supply a feasible method for endogenous microRNA detection in fundamental research and clinical diagnostics.
Keyphrases
- nucleic acid
- living cells
- label free
- fluorescent probe
- single molecule
- real time pcr
- loop mediated isothermal amplification
- quantum dots
- risk assessment
- single cell
- bariatric surgery
- long non coding rna
- mass spectrometry
- signaling pathway
- climate change
- cell therapy
- postmenopausal women
- mesenchymal stem cells
- bone marrow
- fluorescence imaging