Multiplex ratiometric gold nanoprobes based on surface-enhanced Raman scattering enable accurate molecular detection and imaging of bladder cancer
Multiplex ratiometric gold nanoprobes based on surface-enhanced Raman scattering enable accurate molecular detection and imaging of bladder cancer
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基于表面增强拉曼散射的多重比例金纳米探针可实现膀胱癌的精确分子检测和成像
DOI：
10.1007/s12274-021-3902-1
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发表时间：
2021-10
期刊：
影响因子：
9.9
通讯作者：
Fan Jinhai
中科院分区：
文献类型：
--
作者：
Liang Xiao;Zhang Pu;Ma Minghai;Yang Tao;Zhao Xiangwei;Zhang Rui;Jing Minxuan;Song Rundong;Wang Lei;Fan Jinhai
近年来，表面增强拉曼散射（Sers）技术已成功地应用于膀胱肿瘤（BCa）的无创检测。内标法被认为是校正由测量条件和样品干扰引起的信号波动的有效比率测量策略。然而，它仍然是难以检测的目标mRNA定量使用目前的比率Sers纳米传感器。在这项研究中，我们开发了一种基于内部参考的比率Sers检测。将两种携带拉曼报告分子的分子信标（MB）锚定在海胆状Au纳米团簇（AuNCs）上。使用具有六氯荧光素（HEX）的胸苷激酶1（TK 1）MB捕获肿瘤生物标志物TK 1 mRNA，使用具有5（6）-羧基荧光素（FAM）的甘油醛3-磷酸脱氢酶（GAPDH）MB提供内标信号。内参GAPDH MB可反映单细胞内GAPDH mRNA含量的一致性。比值法（I745/I645）能更准确地反映单个细胞中靶mRNA的含量。比率纳米探针具有优异的稳定性（变异系数：0.3%）、高灵敏度（检测限：3.4pM）、高特异性（能够识别单碱基错配）和抗核酶稳定性。值得注意的是，纳米探针可以有效地区分BCa细胞与正常细胞，并且使用比率法很容易描绘单个BCa细胞的轮廓。通过结合不对称聚合酶链反应（PCR）和比率纳米探针，可以很容易地区分低浓度（10− 14 M）的Sers比率（I745/I645）。对BCa患者尿液标本的进一步临床检测证实了其早期无创诊断BCa的潜力，其敏感性为80%，特异性为100%，优于目前尿细胞学方法的上级。
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 (I745/I645) 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 combining asymmetric polymerase chain reaction (PCR) and ratiometric nanoprobes, it was easy to distinguish the SERS ratio (I745/I645) as low concentration as 10−14M. Further clinical detection in urine samples from patients with BCa confirmed its potential for early noninvasive diagnosis of BCa with the sensitivity of 80% and specificity of 100%, which is superior to the current urine cytological method.
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影响因子：
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作者：
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