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血清多肽是癌症诊断信息的重要来源,建立、优化了检测多肽标志物的直接ELISA法,并应用于肝癌血清中的多肽标志物的检测。制备及纯化针对多肽标志物Pep5的单克隆抗体并进行辣根过氧化物酶标记,用其建立检测相应抗原的直接ELISA法。方法线性范围为1.5-20 ng/mL,检测限为1.24 ng/mL;标准品批内及批间CV分别小于3.66%及4.89%,血清样本批内及批间CV分别小于11.69%及18.18%;线性范围内(9、12和15 ng/mL)的回收率分别为98.98%,99.61%和101.58%。应用该方法共检测160例正常血清、104例肝硬化及156例肝癌患者血清,正常组与肝硬化组及肝癌组间差异显著(P<0.001),Pep5诊断肝癌的敏感性和特异性分别为80.8%和96.2%。同时检测94例HCC血清中的AFP和Pep5,AFP检出率为63.8%,Pep5检出率为90.4%,AFP联合Pep5检测时,能将HCC的检出率提高至94.7%。
Serum polypeptide is an important source of cancer diagnostic information, establishing and optimizing the direct ELISA method for detecting polypeptide markers and applying it to the detection of polypeptide markers in the serum of liver cancer. Preparation and purification of monoclonal antibody Pep Peptide Peptide and horseradish peroxidase labeling, with its establishment to detect the corresponding antigen direct ELISA method. The linear range of the method was 1.5-20 ng / mL with the detection limit of 1.24 ng / mL. The intra-assay and inter-assay CV were less than 3.66% and 4.89%, respectively. The intra- and inter-assay CVs were less than 11.69% and 18.18% The recoveries in the linear range (9, 12 and 15 ng / mL) were 98.98%, 99.61% and 101.58%, respectively. The serum of 160 patients with normal liver tissue, 104 patients with cirrhosis and 156 patients with hepatocellular carcinoma was detected by this method. There was significant difference between normal group and cirrhosis group and hepatocellular carcinoma group (P <0.001). The sensitivity and specificity of Pep5 in diagnosing hepatocellular carcinoma were 80.8% and 96.2%. The detection of AFP and Pep5 in 94 cases of HCC at the same time was 63.8% for AFP, 90.4% for Pep5, and 94.7% for AFP combined with Pep5.