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基于上转换纳米颗粒与SYBR Green I之间的荧光共振能量转移(LRET)机制检测血清中的磷脂酶A

Detection of phospholipase A in serum based on LRET mechanism between upconversion nanoparticles and SYBR green I.

作者信息

Luo Dengwang, Zhang Chaoyu, Fang Yao, Shen Yiping, Liang Yuan, Xia Yaokun, Wu Fang, Chen Xiaosong, Liu Jingrong, Chen Jinghua, Li Chunyan, Lan Jianming

机构信息

The School of Pharmacy, Fujian Medical University, Fuzhou, Fujian, 350108, PR China.

Department of Plastic Surgery, The Union Hospital of Fujian Medical University, Fuzhou, Fujian, 350001, PR China.

出版信息

Anal Chim Acta. 2021 Jan 25;1143:37-44. doi: 10.1016/j.aca.2020.11.025. Epub 2020 Nov 24.

DOI:10.1016/j.aca.2020.11.025
PMID:33384128
Abstract

Phospholipase A (PLA) may be a vital biomarker for the prediction and diagnosis of some diseases. Consequently, it is of great significance to quantitatively detect PLA in biologic samples. Herein, on the basis of the principle of luminescence resonance energy transfer (LRET) between upconversion nanoparticles (UCNPs) and SYBR Green I (SG), we proposed a technology for the highly sensitive detection of PLA amount. Therein, as an energy receptor, SG will be quantitatively loaded into liposomes firstly. Then, due to the hydrolysis of liposomes under the catalysis of PLA, SG will be released and inserted into the double-stranded DNA (dsDNA) on the surface of UCNPs, which triggers the LRET because of the shortening of effective spatial distance between UCNPs and SG. Under exciting of NIR light, UCNPs emit luminescence at 476 nm, which makes SG emit fluorescence at 522 nm through LRET. Under optimal conditions, the emission intensity ratio (I/I) increased linearly with the PLA amount in the range of 20 U/L to 400 U/L, and the limit of detection (LOD) reached 15 U/L. Here, after comparing with the clinical standard method, it is found that the biosensor is expected to provide a convenient and sensitive assay for the detection of PLA in actual serum samples. Furthermore, such biosensor can also be used to test the inhibitor of PLA.

摘要

磷脂酶A(PLA)可能是某些疾病预测和诊断的重要生物标志物。因此,定量检测生物样品中的PLA具有重要意义。在此,基于上转换纳米颗粒(UCNPs)与SYBR Green I(SG)之间的发光共振能量转移(LRET)原理,我们提出了一种高灵敏度检测PLA含量的技术。其中,作为能量受体,SG首先会被定量加载到脂质体中。然后,由于PLA催化下脂质体的水解,SG会被释放并插入到UCNPs表面的双链DNA(dsDNA)中,由于UCNPs与SG之间有效空间距离的缩短而触发LRET。在近红外光激发下,UCNPs在476nm处发光,通过LRET使SG在522nm处发出荧光。在最佳条件下发射强度比(I/I)在20 U/L至400 U/L范围内随PLA含量呈线性增加,检测限(LOD)达到15 U/L。在此,与临床标准方法比较后发现,该生物传感器有望为实际血清样品中PLA的检测提供一种便捷且灵敏的检测方法。此外,这种生物传感器还可用于测试PLA的抑制剂。

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