用建立的BA-ELISA检测尿毒症患者血清转铁蛋白的含量

BIOTIN-AVIDIN-ELISA ASSAY OF HUMAN SERUM TRANSFERRIN IN HEALTHY ADULTS AND IN PATIENTS WITH UREMIA

过去检测人血清转铁蛋白(Transferrin,Tf)水平,常用总铁结合力(TIBC法)或环状单向免疫扩散法(SRID)进行。不仅用血量大,且方法繁琐,灵敏度差。为此,我们制备提纯了人血清Tf及兔抗人Tf抗体,建立了生物素-抗生物素蛋白-酶联免疫吸附测定法(BA-ELISA)检测Tf含量的方法,检测限量为0.8ng/ml,检测范围在1～120ng/ml。用于检测的血清样本以1:40000稀释,测得健康人(正常组n=40)血清Tf均值为3.02±0.32mg/ml,尿毒症患者(n=19)血清Tf均值为1.73±0.60mg/ml,明显低于正常组(P<0.001)。本法操作简便,灵敏度高,特异性强,血清量仅需10μl,适用于临床普查及对肾病患者的血清Tf含量的检测。

A highly sensitive assaying method for human transferrin (Tf)biotin^avidin enzyme-linked immunosorbent assay (BA-ELISA) is described. The assaying range of this method was 1-120ng/ml of transferrin and the minimal detectable transferrin concentration was about 0.8 ng/ml. The optimal concentratian for coating antibody and biotinylated antibody as well as the horseradish peroxida.se-avidin conjugate were determined by checkerboard titrations against highly purified transferrin and positive sera. The average level of human serum transferrin waa measured in 40 healthy adults and in 19 patients with uremia, being 3.02±0.32 mg/ml and 1.73 ±0.60 nag/ml, respectively. Ten microliters serum Sample at a dilution of 1:40 000 was used in this assaying method. The average recovery was 96.90%, reflecting a high degree of specificity, sensitivity and reproducibility.