前列腺特异抗原光激发化学发光免疫检测方法的建立

Development of light induced chemiluminescent immunoassay for prostate specific antigen

目的建立前列腺特异抗原(Prostate specific antigen,PSA)光激发化学发光免疫(Light induced chemilumines-cent immunoassay,LICA)定量检测方法。方法用PSA多克隆抗体包被受体微粒,PSA单克隆抗体标记生物素,两者与链霉亲和素包被的供体微粒共同组成检测试剂检测PSA,优化测定条件并对方法进行验证。分别采用本方法与Roche公司电化学发光免疫分析法对82份临床标本进行检测,并进行比较分析。结果本方法的分析灵敏度为0.31 ng/ml;批内变异系数为4.66%～6.75%,批间变异系数为5.68%～8.42%;回收率为95.1%～105.2%。两种方法具有较好的相关性,其R2为0.981 5。结论建立的均相化学发光免疫测定方法能够用于血清PSA的定量测定,且其性能指标符合临床要求。

Objective To develop a light induced chemiluminescent immunoassay(LICA) for quantitative determination of prostate specific antigen(PSA).Methods A LICA kit was developed,which consisted of the receptor particles coated with polyclonal antibody against PSA,the monoclonal antibody against PSA coated with biotin,and the donor particles coated with streptavidin.The condition for determination was optimized,and the developed method was verified.A total of 82 clinical samples were determined by the developed kit and electrochemiluminescent(ECL) immunoassay kit(Roche),and the results were compared.Results The analytical sensitivity of the developed method was 0.31 ng / ml.The intra-and inter-coefficients of variation of determination results were 4.66% ～ 6.75% and 5.68% ～ 8.42% respectively,while the recovery rate was 95.1% ～ 105.2%.The determination result by LICA showed high correlation with that by ECL immunoassay,of which the R2 value was 0.981 5.Conclusion The developed homogeneous chemiluminescent immunoassay method may be used for the quantitative detection of serum PSA,of which the performance meets the clinical requirements.