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目的建立可定量测定人血清中前列腺特异性抗原(PSA)含量的夹心ELISA法,研制PSA EIA检测试剂盒.方法从健康男性精液中提取并纯化PSA,分别免疫Balb/c小鼠和山羊制备特异性单克隆抗体和多克隆抗体 ,并以纯化的PSA为标准品 ,建立可定量测定血清中PSA含量的夹心ELISA法。在此基础上组装PSA EIA试剂盒 ,对该试剂盒的特异性、灵敏度、精密度、正确性和稳定性等多项指标进行评价。应用该试剂盒检测了615份临床标本和部分血清样品 ,并与美国Hybritech公司经FDA批准的Tandem EPSA检测试剂盒进行了比较。结果我们研制的试剂盒的特异性、灵敏度、精密度、正确性和稳定性等多项指标 ,均已达到临床应用要求 ,与国外进口试剂盒的相关性良好 (r=0.983)。结论研制的PSA EIA试剂盒为临床早期诊断前列腺癌提供了有用的工具 ,适合在国内推广应用。
Objective To establish a sandwich enzyme-linked immunosorbent assay (ELISA) for the quantitative determination of prostate-specific antigen (PSA) in human serum and to develop a PSA EIA assay kit.Methods PSA was extracted and purified from healthy male semen, and immunized Balb / c mice and goats respectively Monoclonal antibodies and polyclonal antibodies. The purified PSA was used as a standard to establish a sandwich ELISA method for the quantitative determination of PSA in serum. On this basis, PSA EIA kit was assembled to evaluate the specificity, sensitivity, precision, accuracy and stability of the kit. Using this kit, 615 clinical samples and partial serum samples were tested and compared with the U.S.-based Hybritech’s FDA-approved Tandem EPSA assay kit. Results The specificity, sensitivity, precision, accuracy and stability of the kit we developed have reached the clinical application requirements, and the correlation with the imported kit was good (r = 0.983). Conclusions The developed PSA EIA kit provides a useful tool for the early diagnosis of prostate cancer in clinic and is suitable for popularization and application in China.