肺癌标志物流式微球阵列的分析性能评价

目的 通过探讨检测下限、精密度、特异性、方法学比对和线性范围，评价基于流式细胞仪的肺癌标志物流式微球阵列的分析性能。方法 利用流式细胞仪，测试肺癌标志物流式微球阵列试剂盒检测血清中肺癌标志物癌胚抗原（CEA）、细胞角蛋白19片段（Cyfra21-1）和神经元特异性烯醇化酶（NSE）的检测下限、精密度、特异性和线性范围；以蛋白质印迹（Western blotting）法验证抗体识别抗原的专一性；检测血红蛋白、三酰甘油、胆红素对CEA、Cyfra21-1和NSE检测的干扰作用；通过与电化学发光免疫分析法比对，考察了肺癌标志物流式微球阵列的准确性。结果 CEA、Cyfra21-1和NSE的检测下限分别为1.71、3.97、2.27pg/mL，批内精密度均≤10%，批间精密度均≤15%；特异性结果显示，CEA、Cyfra21-1、NSE的配对抗体能分别专一识别抗原，CEA与同源类似物癌胚抗原相关黏附分子6（CEACAM6）、cyfra21-1与重组人细胞角蛋白18（CK18）、NSE与非神经元特异性烯醇化酶（NNE）无明显交叉反应；三酰甘油、胆红素对血清样本检测无显著干扰作用，500 ng/mL的血红蛋白能够明显干扰Cyfra21-1（P<0.05）和NSE（P<0.05）的检测；流式微球阵列和电化学发光免疫分析的CEA、Cyfra21-1、NSE检测结果的相关系数值分别为0.9842、0.9622、0.982 0；CEA、Cyfra21-1、NSE的线性范围分别为355.76 pg/mL～367.74 ng/mL、87.89 pg/mL～107.8 ng/mL、90.12 pg/mL～86.07 ng/mL。结论 肺癌标志物流式微球阵列的分析性能符合要求。

Analytical capability of flow cytometric bead array for lung cancer markers

Objective To appraise the analytical capability of flow cytometric bead array for lung cancer markers through the tests of limit of detection, relative standard deviation, specificity, methods comparation and linearity rang. Methods The limit of detection, relative standard deviation, specificity and linearity rang in detection of Carcinoembryonic antigen (CEA), cytokeratin 19 (Cyfra21-1) and neuron specific enolase (NSE) in serum were evaluated by flow cytometer. Western blotting method was ultilized to validate the specificity of antibody-antigen recognization.The interference of hemoglobin, three acyl glycerol and bilirubin on the detection of CEA, Cyfra21-1 and NSE was tested. Compared to electrochemiluminescence immunoassay, the relative error for flow cytometric bead array was assessed.Results Flow cytometric bead array demonstrated that the limit of detection was 1.71pg/mL for CEA, 3.97pg/mL for cyfra21-1, and 2.27pg/mL for NSE. The relative standard deviation for intra-assay and inter-assay were below 10% and 15%, respectively. The pair of antibodies can defferentially recognize antigens. The measurement for CEACAM6, CK18, NSE appeared that there was no significant cross-talking reaction. Three acyl glycerol and bilirubin did not significantly interfere with the detection for serum samples. Hemoglobin of 500 ng/mL can significantly interfere with the detection of Cyfra21-1 (P< 0.05) and NSE (P< 0.05). The correlation coefficient between flow cytometric array and electrochemiluminescence immunoassay was 0.9842 for serum CEA, 0.9622 for serum cyfra 21-1 and 0.982 0 for serum NSE. The linearity ranged from 355.76pg/mL to 367.74 ng/mL for CEA, from 87.89 pg/mL to 107.8 ng/mL for cyfra21-1, and from 90.12pg/mL to 86.07 ng/mL for NSE. Conclusion Flow cytometric array for lung cancer markers may be of use in clinical detection.