Luczynska C M, Arruda L K, Platts-Mills T A, Miller J D, Lopez M, Chapman M D
Department of Internal Medicine, University of Virginia, Charlottesville 22908.
J Immunol Methods. 1989 Mar 31;118(2):227-35. doi: 10.1016/0022-1759(89)90010-0.
A two-site monoclonal antibody (Mab) ELISA was developed to measure the Group I allergens from Dermatophagoides spp., Der p I from D. pteronyssinus and Der f I from D. farinae. Species-specific Mabs were used to coat microtiter plates which were then incubated with allergen or house dust extracts. Bound allergen was detected using a biotinylated Mab which recognized a common epitope on both Der p I and Der f I, followed by the addition of streptavidin-peroxidase and ABTS/H2O2 substrate. The assay had low non-specific binding (approximately 0.08 absorbance units) and had a sensitivity of 5 ng/nl for aqueous allergen extracts (equivalent to 0.1 microgram allergen/g dust). 53 dust samples were assayed using the Mab ELISA and an RIA previously described using 125I-labelled Mab. The results showed a very good quantitative correlation between the assays (r = 0.96, p less than 0.001 for Der p I; r = 0.92, P less than 0.001 for Der f I). A further 132 dust samples from a different geographical areas were also assayed by both methods and gave correlation coefficients of 0.90 (P less than 0.001) and 0.86 (P less than 0.001) for Der p I and Der f I, respectively. The Mab ELISA will be useful in epidemiological studies of allergic asthma, both in the assessment of levels of dust mite allergen present in houses and the efficacy of allergen avoidance regimes.
开发了一种双位点单克隆抗体（Mab）酶联免疫吸附测定法（ELISA），用于检测嗜皮螨属中的Ⅰ类变应原，即来自柏氏嗜皮螨的Der p I和来自粉尘螨的Der f I。使用种特异性单克隆抗体包被微量滴定板，然后与变应原或屋尘提取物一起孵育。使用识别Der p I和Der f I上共同表位的生物素化单克隆抗体检测结合的变应原，随后加入链霉亲和素 - 过氧化物酶和ABTS/H₂O₂底物。该测定法具有低非特异性结合（约0.08吸光度单位），对水性变应原提取物的灵敏度为5 ng/nl（相当于0.1微克变应原/克灰尘）。使用Mab ELISA和先前描述的使用¹²⁵I标记单克隆抗体的放射免疫测定法（RIA）对53份灰尘样品进行了检测。结果显示两种测定法之间具有非常好的定量相关性（Der p I的r = 0.96，p < 0.001；Der f I的r = 0.92，P < 0.001）。还通过这两种方法对来自不同地理区域的另外132份灰尘样品进行了检测，Der p I和Der f I的相关系数分别为0.90（P < 0.001）和0.86（P < 0.001）。Mab ELISA在过敏性哮喘的流行病学研究中将会有用，可用于评估房屋中存在的尘螨变应原水平以及变应原回避方案的效果。
