Yasueda H, Akiyama K, Maeda Y, Hayakawa T, Kaneko F, Hasegawa M, Shida T
Clinical Research Center for Allergy and Rheumatology, National Sagamihara Hospital.
Arerugi. 1991 Sep;40(9):1218-25.
A simple enzyme-linked immunosorbent assay (ELISA) has been developed for the quantitation of the major allergens of sugi pollen, Cry j I and of Dermatophagoides mites, Der I (Der p I/Der f I) and Der II (Der p II/Der f II) for use in the in vitro standardization of allergen extracts. Polystyrene microplates coated with a IgG fraction of rabbit antiserum were incubated first with allergen extracts and then with biotinylated antiserum IgG. The bound allergen-biotinylated antibody complex was detected with commercially available streptavidin-enzyme conjugate followed by the addition of colorimetric substrate. The assay was very sensitive (-0.2 ng/ml) and reproducible (CV% = 1.9-13.8%). The ELISA was compared with the radioimmunoassay previously described, and the results showed a very good correlation between the assays (r = 0.967-0.990). The allergen content in three sugi pollen and three house dust extracts measured by the ELISA also demonstrated a good agreement with the relative potency of these extracts as determined by the intradermal skin test. These results indicate that the ELISA could be useful in the standardization of allergen extracts.
已开发出一种简单的酶联免疫吸附测定法（ELISA），用于定量日本柳杉花粉的主要过敏原Cry j I以及粉尘螨的Der I（Der p I/Der f I）和Der II（Der p II/Der f II），以用于过敏原提取物的体外标准化。首先将涂有兔抗血清IgG组分的聚苯乙烯微孔板与过敏原提取物孵育，然后与生物素化抗血清IgG孵育。用市售的链霉亲和素 - 酶偶联物检测结合的过敏原 - 生物素化抗体复合物，随后加入比色底物。该测定法非常灵敏（-0.2 ng/ml）且可重复（CV% = 1.9 - 13.8%）。将该ELISA与先前描述的放射免疫测定法进行比较，结果表明两种测定法之间具有非常好的相关性（r = 0.967 - 0.990）。通过ELISA测量的三种日本柳杉花粉和三种屋尘提取物中的过敏原含量，也与通过皮内皮肤试验确定的这些提取物的相对效价显示出良好的一致性。这些结果表明ELISA可用于过敏原提取物的标准化。
