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目的建立压电石英晶片免疫传感器测定方法检测人血清中胸腺嘧啶脱氧核苷激酶-1(TK1),探索其对常见恶性肿瘤早期诊断的可行性。方法克隆人TK1基因,表达并纯化TK1基因融合蛋白,再制备并纯化TK1单克隆抗体后,用生物素标记TK1单克隆抗体。再选择并利用特殊TK1固化剂和镀金压电石英晶体制备压电石英晶体TK1免疫传感器及抗干扰反应池及屏蔽系统。最后,用此免疫传感器方法检测正常人、不同肿瘤患者血清。结果肝癌血清的TK1水平最高(2·44±0·46)ng/ml,其次为乳腺癌血清(1·16±0·24)ng/ml,二者的TK1水平均明显高于其他恶性肿瘤血清(P<0·001)。肺癌、胃肠癌、生殖系统癌以及其他恶性肿瘤血清间的TK1水平接近(P>0·05),但与良性肿瘤血清TK1存在显著性差异(P<0·001)。此外,良性肿瘤血清的TK1频移值亦显著大于正常人血清(P<0·001)。结论压电石英晶片免疫传感器可作为一种简便灵敏TK1测定新方法用于临床恶性肿瘤普查以及区分肿瘤类型,尤其是肝癌的诊断。
Objective To establish a piezoelectric quartz chip immunosensor assay for the detection of thymidine kinase-1 (TK1) in human serum and explore its feasibility for early diagnosis of common malignant tumors. Methods TK1 gene was cloned and TK1 gene fusion protein was expressed and purified. After TK1 monoclonal antibody was prepared and purified, TK1 monoclonal antibody was labeled with biotin. And then select and use special TK1 curing agent and gold-plated piezoelectric quartz crystal piezoelectric quartz crystal TK1 immune sensor and anti-interference pool and shielding system. Finally, the immunosensor method is used to detect the serum of normal people and different tumor patients. Results The level of TK1 in the serum of hepatocellular carcinoma was the highest (2.44 ± 0.46 ng / ml), followed by the serum of breast cancer (1.16 ± 0.24 ng / ml), the TK1 levels of the two were significantly higher than those of other malignant tumors Serum (P <0.001). The levels of TK1 in lung cancer, gastrointestinal cancer, reproductive system cancer and other malignant tumors were close (P> 0.05), but significantly different from those in benign tumors (P <0.001). In addition, benign tumor serum TK1 frequency shift was significantly greater than normal human serum (P <0.001). Conclusion Piezoelectric quartz wafer immunosensor can be used as a simple and sensitive method for the determination of TK1 in clinical malignant tumor screening and differentiation of tumor types, especially in the diagnosis of liver cancer.