流式荧光免疫法检测多肿瘤标志物的方法学评价

Methodology evaluation of the determination of tumor markers based on flowcytometry fluorescence immunoassay method

目的评价流式荧光免疫法在多肿瘤标志物定量测定中的可靠性。方法用流式荧光免疫法对试剂盒精密度、线性、携带污染率、抗干扰性等指标进行评价,同时将Luminex^TM-100与Bio-Ekon SerozymeⅠ的结果进行比较。结果本方法化学性能的评价表明,批内变异系数（CV）均〈10%,批间CV均〈14%。线性良好（r〉0.99）。携带污染率均〈0.5%。血红蛋白（Hb）、胆红素（Bil）、三酰甘油（TG）对测定结果无显著干扰（P〉0.05）。与Bio-Ekon SerozymeⅠ内分泌定量分析仪及配套试剂相关性良好（r〉0.975）。对506名年龄在24-84岁健康体检人员标本用本方法进行多肿瘤标志物测定,正常参考值分别为甲胎蛋白（AFP）〈15 ng/mL、糖类抗原125（CA125）〈35 U/mL、癌胚抗原（CEA）〈5 ng/mL、糖类抗原199（CA199）〈35 U/mL、前列腺特异抗原（PSA）〈4 ng/mL。结论本方法测定结果准确、可靠且操作简便、快速,适宜于临床检测推广。

Objective To evaluate the reliability of the quantitative determination of tumor markers using reagent kit based on the flowcytometry fluorescence immunoassay method.Methods The precision,linearity,rate of carryover and interference-resistance of the kits were evaluated using the automated analyzer Luminex^TM-100 as determination instrument,and the results were compared with a comparative determination using Bio-Ekon Serozyme Ⅰ system.Results The chemical character evaluation showed that the within-run and between-run coefficients of variation（CV） were less than 10% and 14% respectively.The results demonstrated excellent linearity（r0.99）.The rates of carryover were generally acceptable（0.5%）.The assay had no significant interference by haemoglobin（Hb）,bilirubin（Bil） and triglyceride（TG）（P0.05）.The correlation between the evaluated kits with LuminexTM-100 and Bio-Ekon Serozyme Ⅰ system was excellent（r0.975）.The tumor markers of serum samples from 506 healthy subjects（24 years old to 84 years old） were determined,and the reference result was alpha fetoprotein（AFP）15 ng/mL,carbohydrate antigen 125（CA125）35 U/mL,carcinoembryonic antigen（CEA）5 ng/mL,carbohydrate antigen 199（CA199）35 U/mL and prostate specific antigen（PSA）4 ng/mL.Conclusions The quantitative assay is safe,precise,accurate and easy to operate,and it is worthy to extend for clinical application.