Eye Institute, Eye and ENT Hospital, College of Medicine, Fudan University, Shanghai, China; Shanghai Key Laboratory of Visual Impairment and Restoration, Science and Technology Commission of Shanghai Municipality, Shanghai, China; Key Laboratory of Myopia, Ministry of Health, Shanghai, China.
Department of Laboratory Diagnosis, Changhai Hospital, Second Military Medical University, Shanghai, China.
Biosens Bioelectron. 2018 Apr 15;102:57-62. doi: 10.1016/j.bios.2017.11.017. Epub 2017 Nov 7.
Accurate, fast and sensitive detection of disease-specific protein biomarkers, especially in blood, urine, or other bodily fluids, is an important approach to achieve early disease diagnosis. Platelet-derived growth factor-BB (PDGF-BB), a widely used biomarker, is involved in a substantial number of serious diseases, such as hepatic fibrosis, atherosclerosis, age-related macular degeneration and diabetic eye disease and is often over-expressed in human malignant tumors. Therefore, the development of sensitive and specific detection methods for PDGF-BB is of great importance for the early diagnosis of disease and assessments of patient recovery. In the current study, a biolayer interferometry-based enzyme-linked aptamer sorbent assay (BLI-ELASA) was successfully established for rapid (20-25min), high-throughput (8 or 16 samples) and real-time monitoring of PDGF-BB in clinical samples. The method exhibited a broad detection range from 0.5 to 1000ng/mL of PDGF-BB (good linear range from 0.5 to 10ng/mL), with a low detection limit of 0.08ng/mL. Moreover, BLI-ELASA was applied to the detection of PDGF-BB in spiked serum and urine samples and showed a high degree of selectivity for PDGF-BB, good reproducibility, and stability. We believe that the methodology in this work can be easily adapted to detect other biomolecules in clinical samples, including viruses, pathogens and toxins, in a rapid, sensitive, high-throughput and real-time manner.
准确、快速和敏感地检测疾病特异性蛋白质生物标志物，特别是在血液、尿液或其他体液中，是实现早期疾病诊断的重要方法。血小板衍生生长因子-BB（PDGF-BB）是一种广泛使用的生物标志物，涉及许多严重疾病，如肝纤维化、动脉粥样硬化、年龄相关性黄斑变性和糖尿病眼病，并且经常在人类恶性肿瘤中过度表达。因此，开发用于 PDGF-BB 的敏感和特异检测方法对于疾病的早期诊断和患者恢复情况的评估非常重要。在本研究中，成功建立了基于生物层干涉法的酶联适体吸附测定法（BLI-ELASA），用于快速（20-25min）、高通量（8 或 16 个样本）和实时监测临床样本中的 PDGF-BB。该方法对 PDGF-BB 的检测范围从 0.5 到 1000ng/mL（良好的线性范围为 0.5 到 10ng/mL），检测限低至 0.08ng/mL。此外，BLI-ELASA 被应用于检测加标血清和尿液样本中的 PDGF-BB，对 PDGF-BB 具有高度选择性、良好的重现性和稳定性。我们相信，这项工作中的方法可以很容易地适应于快速、敏感、高通量和实时地检测临床样本中的其他生物分子，包括病毒、病原体和毒素。
