Peng Z, Xu W, Simons F E
Department of Pediatrics and Child Health, University of Manitoba, Winnipeg, Canada.
Ann Allergy Asthma Immunol. 1998 Mar;80(3):274-8. doi: 10.1016/S1081-1206(10)62970-5.
Dermatophagoides farinae (Der f) is a major allergen in Der f extract. Measurement of Der f I-specific IgE has not been commonly used, because of the difficulty in obtaining large amounts of purified Der f 1.
To improve the diagnosis of dust mite allergy, we wanted to develop an ELISA to measure Der f 1-specific IgE in which purified Der f 1 is not required.
Using a monoclonal antibody (mAb) against Der f 1, a mAb-capture ELISA was developed. Microplates coated with the mAb were sequentially incubated with crude Der f extract, serum samples, goat anti-human IgE, and conjugated rabbit anti-goat IgG. A conventional ELISA using purified Der f 1 as capture (Der f 1-ELISA) was developed to compare it with the mAb-capture ELISA.
There was a significant correlation between the two ELISAs (r = 0.89, P < .001) with a sensitivity of 0.8 U/mL. Both the assays were inhibited by the crude Der f extract in a dose-dependent manner. Using the two ELISAs, serum Der f 1-specific IgE was evaluated in 12 allergic patients with positive skin tests to Der f extract, 14 allergic patients with negative skin tests to Der f extract, and 15 healthy subjects. The mean Der f 1-IgE and the positivity rate of Der f I-IgE were higher in the mite-allergic group and lower in the healthy group when the mAb-ELISA was used (P < .01) than when the Der f 1-ELISA was used (P < .05).
Der f 1-specific IgE can be measured by mAb-capture ELISA using crude Der f extract. The mAb-ELISA is more sensitive and specific than the conventional Der f 1-capture ELISA in diagnosing dust mite allergy.
粉尘螨（Der f）是粉尘螨提取物中的主要变应原。由于难以获得大量纯化的Der f 1，Der f 1特异性IgE的检测尚未普遍应用。
为改善尘螨过敏的诊断，我们希望开发一种无需纯化Der f 1即可检测Der f 1特异性IgE的酶联免疫吸附测定（ELISA）。
使用抗Der f 1单克隆抗体（mAb），开发了一种mAb捕获ELISA。将包被有mAb的微孔板依次与粗制粉尘螨提取物、血清样本、山羊抗人IgE和偶联的兔抗山羊IgG孵育。开发了一种以纯化的Der f 1作为捕获物的传统ELISA（Der f 1-ELISA），以与mAb捕获ELISA进行比较。
两种ELISA之间存在显著相关性（r = 0.89，P <.001），灵敏度为0.8 U/mL。两种检测均被粗制粉尘螨提取物以剂量依赖性方式抑制。使用这两种ELISA，对12名对粉尘螨提取物皮肤试验阳性的过敏患者、14名对粉尘螨提取物皮肤试验阴性的过敏患者和15名健康受试者的血清Der f 1特异性IgE进行了评估。使用mAb-ELISA时，螨过敏组的平均Der f 1-IgE和Der f I-IgE阳性率高于健康组（P <.01），而使用Der f 1-ELISA时（P <.05）。
可使用粗制粉尘螨提取物通过mAb捕获ELISA检测Der f 1特异性IgE。在诊断尘螨过敏方面，mAb-ELISA比传统的Der f 1捕获ELISA更敏感、更特异。
