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目的通过建立多种肿瘤标志物联合检测的液相芯片系统,以期达到早期发现、早期诊断肿瘤的目的,提高肿瘤的治疗效果。方法根据Luminex公司所提供的实验流程进行荧光微球表面捕获抗体的包被,并对耦联效率和交叉反应进行鉴定,利用重组人肿瘤标志物蛋白纯品构建多指标联合检测的液相芯片的标准曲线,并与常规检查方法酶联免疫吸附实验(ELISA)进行对比。结果鉴定结果证实抗体包被成功有效,且无交叉反应的发生；但仍应对整个流程进行严格质控,因为耦联过程中的细微差别均可导致微球表面抗体密度的较大变化,并影响后继检测效率；标准曲线构建成功,与传统“金标准”ELISA相比,液相芯片在多指标联合检测潜能、可重复性、置信区间范围、时间及试剂耗费等多方面均有明显优越性。结论成功构建了多种肿瘤标志物联合检测的液相芯片系统,从而为后继大规模血清样本的临床检测和肿瘤筛查莫定了基础。
Objective To establish a liquid-phase microarray system for detection of various tumor markers in order to achieve the purpose of early detection, early diagnosis of the tumor and improvement of the therapeutic effect of the tumor. Methods According to the experimental procedure provided by Luminex Corporation, the surface of fluorescent microspheres was coated with antibody and the coupling efficiency and cross-reaction were identified. The recombinant human tumor marker protein was used to construct multi-index combined liquid-phase chip Standard curve, and compared with the routine test method of enzyme-linked immunosorbent assay (ELISA). Results The results showed that the antibody coating was successfully and effectively performed without cross-reaction. However, strict control of the whole process should be carried out because the slight differences in the coupling process can lead to large changes of the antibody density on the surface of the microsphere and influence Successive detection efficiency; the standard curve was successfully constructed. Compared with the traditional “gold standard” ELISA, the LC chip has obvious advantages in multi-index joint detection potential, repeatability, confidence interval range, time and reagent consumption. Conclusion The successful establishment of a series of liquid-phase microarray system for the detection of tumor markers, which lays the foundation for the clinical detection and tumor screening of the subsequent large-scale serum samples.