Konishi E, Uehara K
Department of Medical Zoology, Kobe University School of Medicine, Japan.
J Med Entomol. 1990 Nov;27(6):993-8. doi: 10.1093/jmedent/27.6.993.
Double-antibody sandwich enzyme-linked immunosorbent assay (ELISA) using a monoclonal antibody as capture and detector antibodies was developed for quantifying antigens of Dermatophagoides farinae Hughes and D. pteronyssinus (Trouessart) mites contained in house dust samples. This monoclonal antibody was directed against mite antigens that were also reactive to immunoglobulin E antibody in all of 10 serum samples obtained from patients allergic to mites. Histological study using fluorescent antibody revealed that the monoclonal antibody was bound to the major part of the D. farinae mite body section including fecal matter and cuticles. The detection limit of the assay system was 0.17 microgram of soluble antigens of both mite species and the antigen amount corresponding to 0.5 mites per microplate well, whether in live or dead mites. This system did not react to antigens of Tyrophagus putrescentiae (Schrank) and Cheyletus malaccensis Oudemans. Slight inhibition of less than or equal to 21.3% was observed with nonspecific substances contained in house dust, such as wool, cotton, human dander, human hair, soil, and biscuit, but no direct ELISA reactions were obtained with any of these materials. In 49 house dust samples, ELISA was significantly correlated with the conventional microscopic observation method.
开发了一种双抗体夹心酶联免疫吸附测定（ELISA）方法，该方法使用单克隆抗体作为捕获抗体和检测抗体，用于定量室内灰尘样本中所含的粉尘螨（Dermatophagoides farinae Hughes）和屋尘螨（D. pteronyssinus (Trouessart)）的抗原。这种单克隆抗体针对的螨抗原，对从螨过敏患者获得的10份血清样本中的所有样本，也与免疫球蛋白E抗体发生反应。使用荧光抗体的组织学研究表明，单克隆抗体与粉尘螨螨体切片的主要部分结合，包括粪便和角质层。该检测系统的检测限为两种螨类可溶性抗原的0.17微克，以及每微孔板孔对应于0.5只螨的抗原量，无论螨是活的还是死的。该系统对腐食酪螨（Tyrophagus putrescentiae (Schrank)）和马六甲肉食螨（Cheyletus malaccensis Oudemans）的抗原无反应。对室内灰尘中含有的羊毛、棉花、人皮屑、人发、土壤和饼干等非特异性物质观察到小于或等于21.3%的轻微抑制，但这些材料均未获得直接ELISA反应。在49份室内灰尘样本中，ELISA与传统显微镜观察方法显著相关。
