Cheng Yu-Hong, Tang Hao, Yu Ru-Qin, Jiang Jian-Hui
State Key Laboratory of Chemo/Biosensing and Chemometrics, College of Chemistry and Chemical Engineering, Hunan University, Changsha 410082, P. R. China.
Analyst. 2020 Jul 13;145(14):4860-4866. doi: 10.1039/d0an00656d.
We report a novel DNA-programmed plasmonic enzyme-linked immunosorbent assay (ELISA) for the ultrasensitive detection of protein biomarkers with the naked eye. The DNA-programmed plasmonic assay was based on two enzyme-free and isothermal nucleic acid amplification methods: hybridization chain reaction (HCR) and catalyzed hairpin assembly (CHA). In this study, a biotin-labeled DNA probe was utilized insteand of an enzyme-label probe in well-developed ELISA method. The biotin-labeled DNA probe was able to trigger the HCR and CHA processes, and the products could hybridize with DNA-modified gold nanoparticles (AuNPs) to induce the aggregation of the AuNPs and a color change in the solution. The developed method was able to detect as low as 1 pg mL-1 PSA target with the naked eye. Clinical serum samples demonstrated satisfactory results, indicating that the method is useful for early diagnostics and monitoring curative effects after a medical treatment. The developed method presents a simple and portable platform for ultrasensitive protein detection and has potential for point-of-care (POC) diagnostics in less developed areas.
我们报道了一种新型的DNA编程等离子体酶联免疫吸附测定法(ELISA),用于肉眼超灵敏检测蛋白质生物标志物。该DNA编程等离子体测定法基于两种无酶等温核酸扩增方法:杂交链式反应(HCR)和催化发夹组装(CHA)。在本研究中,在成熟的ELISA方法中使用生物素标记的DNA探针代替酶标记探针。生物素标记的DNA探针能够触发HCR和CHA过程,其产物可与DNA修饰的金纳米颗粒(AuNPs)杂交,诱导AuNPs聚集并使溶液颜色发生变化。所开发的方法能够肉眼检测低至1 pg mL-1的PSA靶标。临床血清样本显示出令人满意的结果,表明该方法可用于早期诊断和治疗后疗效监测。所开发的方法为超灵敏蛋白质检测提供了一个简单便携的平台,在欠发达地区具有即时检测(POC)诊断的潜力。