Department of Biomedical Information Center, Beijing You'an Hospital, Capital Medical University, Beijing, PR China.
Beijing Qingyuan Bio-Technologies Inc, Beijing, PR China.
Clin Chim Acta. 2018 Jul;482:209-214. doi: 10.1016/j.cca.2018.03.039. Epub 2018 Apr 6.
Development of a new method to simultaneously detect Alpha-fetoprotein (AFP) and Golgi protein 73 (GP73) from peripheral blood.
Anti human AFP and GP73 monoclonal antibodies was used to develop a sandwich immunity reaction using xMAP technology for the simultaneous detection of plasma AFP and GP73. The assay evaluated the sensitivity, cross reactivity, range of detection, precision, recovery and linearity dilution effect. The assay utilized plasma samples and compared its performance with commercially available Enzyme Linked Immunosorbent Assay (ELISA) kits.
The assay was successful in detecting AFP and GP73 simultaneously. Validation experiments demonstrated the limit of detection was AFP 0.006 μg/l and GP73 0.482 μg/l. The functional sensitivity was AFP 0.010 μg/l and GP73 0.640 μg/l. The range of detection was AFP 0.01-50 μg/l and GP73 0.64-100 μg/l. No cross reactivity was observed. The intra- and inter-assay variation for AFP was 0.19-3.46% and 3.1-5.8% and for GP73 was 1.5-3.2% and 1.1-7.6% respectively. The recovery for AFP was 96-106% and GP73 was 89-110%. 80 clinical plasma samples from healthy controls, and patients with liver cirrhosis and Hepatocellular Carcinoma (HCC) were evaluated. For healthy controls (n = 25), the AFP was 2.40 (1.55, 3.30) μg/l and GP73 was 42.60 (39.10, 57.40) μg/l. For patients with liver cirrhosis (n = 19), the AFP was 2.60 (1.70, 4.20) μg/l and GP73 was 136.10 (92.10, 261.70) μg/l, and for HCC patients (n = 36), the AFP was 13.65 (3.35, 158.88) μg/l and GP73 was 186.25 (96.73, 262.03) μg/l. The new assay demonstrated a good correlation with commercially available ELISA kits (correlation coefficients (r) were 0.997 (AFP, p < 0.001) and 0.959 (GP73, p < 0.001).
The method demonstrated a sensitive, effective and accurate method for the simultaneous detection of AFP and GP73, and could be used clinically for routine detection and monitoring of patients with chronic hepatitis B.
开发一种新方法，同时从外周血中检测甲胎蛋白（AFP）和高尔基糖蛋白 73（GP73）。
使用抗人 AFP 和 GP73 单克隆抗体，利用 xMAP 技术开发夹心免疫反应，同时检测血浆 AFP 和 GP73。该检测评估了灵敏度、交叉反应性、检测范围、精密度、回收率和线性稀释效应。该检测使用了血浆样本，并将其性能与市售的酶联免疫吸附测定（ELISA）试剂盒进行了比较。
该检测成功地同时检测 AFP 和 GP73。验证实验表明，检测限为 AFP 0.006μg/l 和 GP73 0.482μg/l。功能灵敏度为 AFP 0.010μg/l 和 GP73 0.640μg/l。检测范围为 AFP 0.01-50μg/l 和 GP73 0.64-100μg/l。未观察到交叉反应。AFP 的批内和批间变异分别为 0.19-3.46%和 3.1-5.8%，GP73 分别为 1.5-3.2%和 1.1-7.6%。AFP 的回收率为 96-106%，GP73 为 89-110%。评估了 80 份来自健康对照者和肝硬化及肝细胞癌（HCC）患者的临床血浆样本。对于健康对照组（n=25），AFP 为 2.40（1.55，3.30）μg/l，GP73 为 42.60（39.10，57.40）μg/l。对于肝硬化患者（n=19），AFP 为 2.60（1.70，4.20）μg/l，GP73 为 136.10（92.10，261.70）μg/l，对于 HCC 患者（n=36），AFP 为 13.65（3.35，158.88）μg/l，GP73 为 186.25（96.73，262.03）μg/l。新检测方法与市售 ELISA 试剂盒具有良好的相关性（相关系数（r）分别为 0.997（AFP，p<0.001）和 0.959（GP73，p<0.001）。
该方法显示出对 AFP 和 GP73 的检测具有灵敏、有效和准确的特点，可用于临床常规检测和监测慢性乙型肝炎患者。
