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目的研制高灵敏度的质量放大压电石英晶体免疫传感器用于人血清中癌症标志物胸苷激酶1(hTK1)的检测。方法将半胱氨自组装膜用戊二醛活化,共价固定hTK1抗原,通过和待测抗原竞争与一定浓度的酶标抗体反应,应用4-氯-1萘酚和H2O2质量放大体系,测得样品中待测抗原的浓度。结果hTK1的浓度在0.1~10 ng/m l范围内,与传感器频率的变化之间呈良好的线性关系。应用此方法对患有不同癌症病人血清进行测定,发现正常血清与癌症血清的频移值存在显着差异。结论基于质量放大原理的压电石英晶体免疫传感器可定量测定血清中的ng水平的hTK1抗原,有望用于癌症的早期诊断和临床检测。
Objective To develop a high-sensitivity massively-amplified piezoelectric quartz crystal immunosensor for the detection of human cancer marker thymidine kinase 1 (hTK1). Methods The cysteine self-assembled membrane was activated with glutaraldehyde, and the hTK1 antigen was covalently immobilized. By competing with the antigen to be tested and reacting with a certain concentration of enzyme-labeled antibody, 4-chloro-1-naphthol and H2O2 mass- The concentration of the antigen to be tested in the sample. Results The concentration of hTK1 was in the range of 0.1 ~ 10 ng / ml, which showed a good linear relationship with the change of sensor frequency. The application of this method in patients with different cancer serum was measured and found that there is a significant difference between the normal serum and cancer serum frequency shift. Conclusion Piezoelectric quartz crystal immunosensor based on mass amplification principle can quantitatively determine ng level of hTK1 antigen in serum and is expected to be used in the early diagnosis and clinical detection of cancer.