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 μL) 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.

中文翻译：

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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测定。

蛋白质生物标记物已广泛用于疾病诊断，但是集中实验室中目前使用的检测方法主要基于酶联免疫吸附测定（ELISA）衍生的夹心型免疫测定，例如化学发光或电化学发光免疫测定，这些方法检测时间长且繁琐的乐器。对于蛋白质生物标志物的即时检测（POC），已经制造了各种用于侧向流免疫测定（LFIA）的测试条，但是它们对原始样品的检测灵敏度和能力受到限制。在这项研究中，建立了增强的离心辅助侧向流免疫测定（ECLFIA），以比LFIA更高的灵敏度快速检测全血中的蛋白质生物标志物。通过将硝酸纤维素膜插入离心盘，在ECLFIA的离心平台上启用了全自动操作，包括样品制备，主动侧流驱动，清洗和信号放大，而这在常规LFIA中几乎是无法执行的。在一滴血（20μL）中检测人类前列腺特异性抗原（PSA）的整个过程可以在15分钟内完成。我们的ECLFIA系统的检出限为0.028 ng mL^-1，与LFIA相比，提高了21.4倍。此外，该系统用于检测34个临床样品中的PSA。将结果与使用医院中使用的商用仪器测量的结果进行比较，并获得了良好的相关系数0.986，证明了该ECLFIA系统的实用性。综上所述，本研究建立的ECLFIA系统可以作为蛋白质生物标志物POC检测的有效工具，在灵敏度，简便性和速度方面具有综合优势。