An enhanced centrifugation-assisted lateral flow immunoassay for the point-of-care detection of protein biomarkers
An enhanced centrifugation-assisted lateral flow immunoassay for the point-of-care detection of protein biomarkers
复制标题
用于蛋白质生物标志物即时检测的增强型离心辅助侧流免疫分析
DOI：
10.1039/d0lc00518e
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发表时间：
2020-08-07
期刊：
影响因子：
6.1
通讯作者：
Xu, Youchun
中科院分区：
文献类型：
--
作者：
Shen, Minjie;Li, Nan;Xu, Youchun
蛋白质生物标志物被广泛用于疾病诊断，但目前集中式实验室使用的检测方法主要是基于酶联免疫吸附分析(ELISA)衍生的夹心式免疫分析，如化学发光免疫分析或电化学发光免疫分析，检测时间长，仪器繁琐。对于蛋白质生物标志物的POC检测，已经制造了用于侧向流动免疫分析(LFIA)的各种试条，但它们对原始样品的检测灵敏度和能力有限。在这项研究中，建立了一种增强型离心机辅助侧向流动免疫分析(ECLFIA)来快速检测全血中的蛋白质生物标志物，比LFIA具有更高的灵敏度。通过将硝化纤维素膜插入离心盘，在ECLFIA的离心台上实现了全自动操作，包括样品准备、主动侧向流动激励、洗涤和信号放大，这些操作在传统的LFIA中几乎无法完成。检测一滴血(20亩L)中的人前列腺特异性抗原(PSA)的整个过程可以在15分钟内完成。该系统的检出限为0.028 ngmL(-1)，比LFIA提高了21.4倍。并利用该系统对34份临床标本进行了PSA检测。将结果与医院使用的商用仪器的测量结果进行了比较，相关系数为0.986，证明了该系统的实用性。综上所述，本研究建立的ECLFIA体系具有灵敏、简便、快速的综合优势，可作为蛋白质生物标志物POC检测的有效工具。
Protein biomarkers are widely used for disease diagnosis, but the current detection methods utilized in centralized laboratories are mainly based on enzyme-linked immunosorbent assay (ELISA)-derived sandwich-type immunoassays such as chemiluminescent or electrochemiluminescent immunoassays, which suffer from long detection times and cumbersome instruments. For the point-of-care (POC) detection of protein biomarkers, various test strips for lateral flow immunoassay (LFIA) have been manufactured, but their detection sensitivities and capabilities for raw samples are limited. In this study, an enhanced centrifugation-assisted lateral flow immunoassay (ECLFIA) was established to rapidly detect protein biomarkers in whole blood with a higher sensitivity than LFIA. By inserting a nitrocellulose membrane into a centrifugal disc, fully automated operations, including sample preparation, active lateral flow actuation, washing, and signal amplification, which could hardly be performed in conventional LFIA, were enabled on the centrifugal platform for ECLFIA. The entire process for detecting human prostate specific antigen (PSA) in a drop of blood (20 mu L) could be completed in 15 min. The limit of detection for our ECLFIA system was 0.028 ng mL(-1), showing a 21.4-fold improvement compared to that of LFIA. Moreover, this system was utilized to detect PSA in 34 clinical samples. The results were compared to those measured using a commercial instrument used in the hospital, and a good correlation coefficient of 0.986 was obtained, demonstrating the practicality of this ECLFIA system. In summary, the ECLFIA system established in this study can be an efficient tool for the POC detection of protein biomarkers with comprehensive advantages in sensitivity, simplicity and speed.