Multiplex ratiometric gold nanoprobes based on surface-enhanced Raman scattering enable accurate molecular detection and imaging of bladder cancer

作者：Xiao Liang, Pu Zhang, Minghai Ma, Tao Yang, Xiangwei Zhao, Rui Zhang, Minxuan Jing, Rundong Song, Lei Wang, Jinhai Fan · 发表于：Nano Research · 年份：2021 · DOI：10.1007/s12274-021-3902-1 · 被引用次数：24 · 研究领域：Bladder and Urothelial Cancer Treatments

Recently, surface-enhanced Raman scattering (SERS) has been successfully used in the non-invasive detection of bladder tumor (BCa). The internal standard method was considered as an effective ratiometric strategy for calibrating signal fluctuation originated from the interference of measurement conditions and samples. However, it is still difficult to detect the target mRNA quantitatively using the current ratiometric SERS nanosensors. In this study, we developed an internal reference based ratiometric SERS assay. Two kinds of molecular beacons (MB) carrying Raman reporter molecules were anchored on sea-urchin-like Au nanoclusters (AuNCs). Thymidine kinase1 (TK1) MBs with hexachlorofluorescein (HEX) were used to capture tumor biomarker TK1 mRNA, and glyceraldehyde 3-phosphate dehydrogenase (GAPDH) MBs with 5(6)-carboxyfluorescein (FAM) were used to offer internal standard signals. The internal reference GAPDH MB can reflect the consistent content of the GAPDH mRNA in single cells. The ratiometric method ( I 745 / I 645 ) can more accurately reflect the content of target mRNA in single cells. The ratiometric nanoprobes had excellent stability (coefficient of variation: 0.3%), high sensitivity (detection limit: 3.4 pM), high specificity (capable of single-base mismatch recognition) and ribozyme-resistant stability. Notably, the nanoprobes can effectively distinguish BCa cells from normal cells, and it was easy to contour the single BCa cell using the ratiometric method. By comb...