Most cancers developed an elevation of the level of at least two markers associated with their incidence. Simultaneous detection of multi-tumor markers associated with a particular type of cancer plays an important role in cancer diagnostic. Here, a multianalyte immunoassay chip for simple and sensitive detection of tumor markers with chemiluminescent and colorimetric methods was proposed, in which carcinoembryonic antigen (CEA) and carbohydrate antigen (CA19-9) that associated with colorectal cancer were detected as model. The immunoassay chip was fabricated by co-immobilization of CEA/CA19-9 antibody on a glass slide with γ-glycidoxypropyltrimethoxysilane as linkage. Through sandwiched immunoreactions, CEA, CA19-9, and their corresponding enzyme tracers, alkaline phosphatase-labeled anti-CEA and horseradish peroxidase-labeled anti-CA19-9, were introduced on the chip. Then, they were sequentially detected by chemiluminescent method in the range of 0.5–80 μg/L and 0.5–80 kU/L with the detection limits of 0.41 μg/L and 0.36 kU/L at 3σ for CEA and CA19-9, respectively. They could also be detected by colorimetric method in the range of 1–200 μg/L and 5–200 kU/L with the detection limits of 0.25 μg/L and 1.25 kU/L at 3σ for CEA and CA19-9, respectively. All these results demonstrated that the present work provided a promising analytical method for tumor markers’ analysis with the advantages of simple analytical procedure, small sample volume and lower cost, which made the proposed method potential for high-throughput detection.
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机译：大多数癌症的发病率至少升高了两种标记物的水平。与特定类型癌症相关的多肿瘤标志物的同时检测在癌症诊断中起着重要作用。在这里，提出了一种用于化学发光和比色法简单，灵敏地检测肿瘤标志物的多分析物免疫测定芯片，其中检测与大肠癌相关的癌胚抗原（CEA）和糖类抗原（CA19-9）。通过将CEA / CA19-9抗体以γ-环氧丙氧基丙基三甲氧基硅烷为键共固定在载玻片上来制备免疫测定芯片。通过夹心的免疫反应，将CEA，CA19-9及其相应的酶示踪剂，碱性磷酸酶标记的抗CEA和辣根过氧化物酶标记的抗CA19-9引入了芯片。然后，通过化学发光法依次检测它们的浓度范围为0.5–80μg/ L和0.5–80 kU / L，CEA和CA19-9在3σ时的检出限分别为0.41μg/ L和0.36 kU / L。 。还可以通过比色法在1–200μg/ L和5–200 kU / L的范围内进行检测，CEA和CA19-9在3σ时的检出限分别为0.25μg/ L和1.25 kU / L。所有这些结果表明，目前的工作为分析肿瘤标志物提供了一种有希望的分析方法，该方法具有分析过程简单，样品量少，成本低等优点，为高通量检测提供了可能。
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