Ali M, Ramanarayanan M, Connell J T, Nalebuff D J, Fayemi A O, Mesa-Tejada R
Ann Allergy. 1979 Apr;42(4):231-5.
A solid-phase enzyme immunoassay for allergen-specific IgE antibodies in serum is descirbed. In this technique these antibodies are allowed to bind to the allergen previously adsorbed to the wells of polystyrene microtiter plates. After a washing step the tubes are incubated with rabbit antihuman IgE labelled with horseradish peroxidase. Following a second washing step, the enzyme bound to the tubes is assayed spectrophotometrically using O-phenylene diamine as the substrate. The standard curves obtained with this method and with the RAST technique are illustrated. For the assay of serum antiragweed IgE antibodies, concordance between the results obtained with the RAST test and this immunoperoxidase assay was observed in 85 or 95 (90%) patients with symptoms of ragweed hayfever. The coefficients of variation ranged from 4.4% (2SD +/- .010) TO 14% (2SD +/- .008). The advantages of using peroxidase as the enzymatic marker for the assay of allergen-specific IgE are discussed.
本文描述了一种用于检测血清中变应原特异性IgE抗体的固相酶免疫测定法。在该技术中,这些抗体可与预先吸附在聚苯乙烯微量滴定板孔中的变应原结合。经过洗涤步骤后,将试管与用辣根过氧化物酶标记的兔抗人IgE一起温育。经过第二次洗涤步骤后,使用邻苯二胺作为底物,通过分光光度法测定结合在试管上的酶。文中展示了用该方法和放射变应原吸附试验(RAST)技术获得的标准曲线。对于血清抗豚草IgE抗体的检测,在85或95例(90%)有豚草花粉症症状的患者中,观察到RAST试验结果与这种免疫过氧化物酶测定法结果之间具有一致性。变异系数范围为4.4%(2SD±0.010)至14%(2SD±0.008)。文中还讨论了使用过氧化物酶作为酶标记物检测变应原特异性IgE的优点。