前列腺特异性抗原EIA试剂盒的研制及应用

Establishment and clinical application of a ELISA method for the quantitative detection of prostate specific antigen

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。

Aim To establish a sandwich ELISA method for the quantitative detection of prostate specific antigen and to develop the PSA EIA kit. Methods PSA was isolated and purified from human seminal plasma and then the mouse monoclonal and goat polyclonal anti PSA antibodies were raised by immunizing Balb/c mice and goat with the purified PSA. The purified PSA could be used as the standard regent SD a sandwich PSA ELISA method was established by using tae two kinds of antibodies.The specificity,sensitivity,precision, accuracy and stability of this kit were evaluated. Furthermore,the PSA EIA kit was compared with Tendem E PSA assay kit produced by Hybritech Inc and approved by FDA of USA. Results The specificity,sensitivity,precision,accuracy and stabiliity of this kit can meet the requirements of clinical PSA test. Compared with imported kits, its correlative coefficient reached 0.983. Conclusion The established PSA EIA kit provides a new useful tool for the early identification of prostate cancer and is suitable to be used in China.