Zhang Wei, Chen Junlei, Li Xinxia, Wang Yuwen, Li Jiutong
Pharmacy Academy of Xinjiang Medical University, Urumqi, 830054, China.
Chemistry and Chemical Engineering Academy of Xinjiang Normal University, Urumqi, 830054, China.
J Med Biochem. 2019 Mar 3;38(2):172-180. doi: 10.2478/jomb-2018-0017. eCollection 2019 Apr.
To establish a fast and simple quantitative method for detection of heart-type fatty acid-binding protein (H-FABP) in serum based on a background fluorescence quenching immunochromatographic assay.
A detection card based on the double-antibody sandwich double-antibody method with background fluorescence quenching was developed for quantitative measurement of H-FABP in serum. The optimal concentrations of control for coating the test and control lines were determined as well as the concentrations of gold-labeled antibodies used in preparing the detection system. The detection method for H-FABP in serum was established and validated using real-world clinical samples.
The optimal concentrations of labeling antibody and coating antibody were 5.0 μg/mL and 1.0 mg/mL, respectively. The test card had a sensitivity of 1.15 ng/mL over a linear concentration range of 0-100 ng/mL. Based on three batches prepared for testing the card, the relative standard deviation (RSD) within batches was less than 15% without a significant difference (P=0.942). The detection method was tested against common interfering substances in serum, such as bilirubin, triglyceride and serum anticoagulants ethylenediamine tetraacetic acid (EDTA), heparin, and sodium citrate, and no significant cross-reaction was detected. The test method was further validated with 50 clinical serum samples, and the test results were comparable with standard reference detection methods with good correlation (R=0.95).
Our study presents a new method with strong specificity and sensitivity for the detection of H-FABP in serum, which could promote H-FABP detection in a broad range of applications.
基于背景荧光猝灭免疫层析法建立一种快速、简便的血清中心型脂肪酸结合蛋白（H-FABP）定量检测方法。
研制一种基于背景荧光猝灭双抗体夹心法的检测卡，用于血清中H-FABP的定量检测。确定包被检测线和对照线的最佳质控浓度以及制备检测系统所用金标抗体的浓度。使用实际临床样本建立并验证血清中H-FABP的检测方法。
标记抗体和包被抗体的最佳浓度分别为5.0μg/mL和1.0mg/mL。该检测卡的灵敏度为1.15ng/mL，线性浓度范围为0-100ng/mL。基于制备的三批检测卡进行检测，批内相对标准偏差（RSD）小于15%，差异无统计学意义（P=0.942）。该检测方法针对血清中常见干扰物质如胆红素、甘油三酯以及血清抗凝剂乙二胺四乙酸（EDTA）、肝素和柠檬酸钠进行检测，未检测到明显交叉反应。用50份临床血清样本进一步验证该检测方法，检测结果与标准参考检测方法具有可比性，相关性良好（R=0.95）。
本研究提出一种检测血清中H-FABP的新方法，具有较强的特异性和灵敏度，可促进H-FABP在广泛应用中的检测。
