目的 通過探討檢測(cè)下限、精密度、特異性、方法學(xué)比對(duì)和線性范圍，評(píng)價(jià)基于流式細(xì)胞儀的肺癌標(biāo)志物流式微球陣列的分析性能。方法 利用流式細(xì)胞儀，測(cè)試肺癌標(biāo)志物流式微球陣列試劑盒檢測(cè)血清中肺癌標(biāo)志物癌胚抗原（CEA）、細(xì)胞角蛋白19片段（Cyfra21-1）和神經(jīng)元特異性烯醇化酶（NSE）的檢測(cè)下限、精密度、特異性和線性范圍；以蛋白質(zhì)印跡（Western blotting）法驗(yàn)證抗體識(shí)別抗原的專一性；檢測(cè)血紅蛋白、三酰甘油、膽紅素對(duì)CEA、Cyfra21-1和NSE檢測(cè)的干擾作用；通過與電化學(xué)發(fā)光免疫分析法比對(duì)，考察了肺癌標(biāo)志物流式微球陣列的準(zhǔn)確性。結(jié)果 CEA、Cyfra21-1和NSE的檢測(cè)下限分別為1.71、3.97、2.27pg/mL，批內(nèi)精密度均≤10%，批間精密度均≤15%；特異性結(jié)果顯示，CEA、Cyfra21-1、NSE的配對(duì)抗體能分別專一識(shí)別抗原，CEA與同源類似物癌胚抗原相關(guān)黏附分子6（CEACAM6）、cyfra21-1與重組人細(xì)胞角蛋白18（CK18）、NSE與非神經(jīng)元特異性烯醇化酶（NNE）無明顯交叉反應(yīng)；三酰甘油、膽紅素對(duì)血清樣本檢測(cè)無顯著干擾作用，500 ng/mL的血紅蛋白能夠明顯干擾Cyfra21-1（P<0.05）和NSE（P<0.05）的檢測(cè)；流式微球陣列和電化學(xué)發(fā)光免疫分析的CEA、Cyfra21-1、NSE檢測(cè)結(jié)果的相關(guān)系數(shù)值分別為0.9842、0.9622、0.982 0；CEA、Cyfra21-1、NSE的線性范圍分別為355.76 pg/mL～367.74 ng/mL、87.89 pg/mL～107.8 ng/mL、90.12 pg/mL～86.07 ng/mL。結(jié)論 肺癌標(biāo)志物流式微球陣列的分析性能符合要求。

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.

國家自然科學(xué)基金（81171413）