Department of Clinical Laboratory, the Affiliated Drum Tower Hospital of Nanjing University Medical School, Nanjing 210008, China.
Department of Clinical Laboratory, the Second Affiliated Hospital, Nanjing Medical University, Nanjing 210011, China.
Sci Rep. 2017 Jan 5;7:40090. doi: 10.1038/srep40090.
Golgi protein 73 (GP73) is a novel and potential marker for diagnosing hepatocellular carcinoma (HCC) that has been found to be abnormally elevated in liver disease. A latex particle-enhanced turbidimetric immunoassay (LTIA) was recently introduced and licensed for application in a variety of automated clinical chemistry analyzers. However, no studies have reported sufficient data on analytical performance of this method when using 3 monoclonal antibodies for GP73 measurement. The experimental conditions were firstly optimized and range of linearity, diagnostic potential, clinical relevance were compared with the LTIA based on polyclonal antibodies and ELISA. Dilution tests for the LTIA using 3 monoclonal antibodies produced a calibration curve from 10 to 350 ng/mL while the polyclonal antibodies produced the curve from 20 to 320 ng/mL. The detection limit was achieved at 1.82 ng/mL concentration. Within-run CV was obtained in the range of 1.5-2.9% and ROC curves indicated sensitivity and specificity of the LTIA based on 3 monoclonal antibodies were 96.7% and 93.3%, respectively, higher than for the polyclonal antibodies (94.6% and 72.4%) and ELISA (70.0% and 83.3%). Therefore, the LTIA assay based on 3 monoclonal antibodies is thus applicable in quantification of GP73 concentration in automated biochemistry analyzers.
高尔基体蛋白 73（GP73）是一种新型的、有潜力的肝细胞癌（HCC）诊断标志物，在肝脏疾病中发现其异常升高。最近推出了一种胶乳粒子增强比浊免疫测定法（LTIA），并获得许可应用于各种自动化临床化学分析仪。然而，目前尚无研究报道使用 3 种单克隆抗体测量 GP73 时该方法的分析性能数据是否充足。我们首先优化了实验条件，并与基于多克隆抗体和 ELISA 的 LTIA 比较了线性范围、诊断潜能和临床相关性。使用 3 种单克隆抗体进行 LTIA 的稀释试验可产生 10 至 350ng/mL 的校准曲线，而多克隆抗体则可产生 20 至 320ng/mL 的曲线。检测限达到 1.82ng/mL 浓度。批内 CV 在 1.5-2.9%范围内，ROC 曲线表明基于 3 种单克隆抗体的 LTIA 的灵敏度和特异性分别为 96.7%和 93.3%，高于多克隆抗体（94.6%和 72.4%）和 ELISA（70.0%和 83.3%）。因此，基于 3 种单克隆抗体的 LTIA 测定法适用于自动化生化分析仪中 GP73 浓度的定量。
