Barnard G J, Kim J B, Brockelbank J L, Collins W P, Gaier B, Kohen F
Clin Chem. 1984 Apr;30(4):538-41.
We have developed immunoassays, monitored by the detection of chemiluminescence, for measuring choriogonadotropin in human urine. These methods involve the use of derivatives of isoluminol and include: (a) a labeled antigen with a second antibody covalently linked to polyacrylamide beads as the solid-phase reagent (i.e., solid-phase chemiluminescence immunoassay); and (b) an excess concentration of a specific antibody passively adsorbed onto the walls of polystyrene tubes and a labeled antibody of different specificity (i.e., two-site immunochemiluminometric assay). After the respective binding reactions, the solutions are aspirated, the antigen- or antibody-bound fractions are washed twice with 500 microL of buffer, sodium hydroxide (2 mol/L; 200 microL) is added, and the mixture is incubated for 60 min at 60 degrees C. After cooling, the label is oxidized with microperoxidase/hydrogen peroxide and the resulting chemiluminescence signal is measured for 10 s. We have evaluated the methods in terms of their sensitivity, precision, and clinical utility, and we compare results with values obtained by radioimmunoassay.
我们开发了通过检测化学发光进行监测的免疫测定法,用于测量人尿中的绒毛膜促性腺激素。这些方法涉及使用异鲁米诺衍生物,包括:(a) 一种标记抗原,其与共价连接到聚丙烯酰胺珠上的第二抗体作为固相试剂(即固相化学发光免疫测定法);以及 (b) 过量浓度的特异性抗体被动吸附在聚苯乙烯管的壁上,以及具有不同特异性的标记抗体(即两点免疫化学发光测定法)。在各自的结合反应后,吸出溶液,将抗原或抗体结合部分用500微升缓冲液洗涤两次,加入氢氧化钠(2摩尔/升;200微升),并将混合物在60℃下孵育60分钟。冷却后,用微过氧化物酶/过氧化氢氧化标记物,并测量10秒产生的化学发光信号。我们从灵敏度、精密度和临床实用性方面评估了这些方法,并将结果与放射免疫测定法获得的值进行比较。