摘要
目的对粉尘螨（Dermatophagoides farinae）变应原进行分离和鉴定其特异性变应原组分。方法采用ELISA检测粉尘螨变应原的生物活性。按常规方法制备粉尘螨浸出液，经SDS-PAGE分离，测定各组分的相对分子质量（MT）；同时用对螨过敏的60例病人混合血清（建立血清库）作探针进行Westernblot，鉴定其特异性变应原组分。结果SDS-PAGE显示粉尘螨有15条蛋白带，MT在14×10^3～109×10^3之间，其中主带有9条，MT分别为109×10^3、100×10^3、86×10^3、62×10^3、56×10^3、36×10^3、28×10^3、19×10^3、14×10^3；Westernblot结果表明，浸出液中共有5条致敏条带，其MT分别为109×10^3、100×10^3、36×10^3、28×10^3、14×10^3；经ELISA检测粉尘螨浸出液具有生物活性。结论粉尘螨的特异性变应原有5条，分别为109×10^3、100×10^3、36×10^3、28×10^3、14×10^3．其浸出液有稳定的生物活性。该研究为开发适合我国人群的粉尘螨标准化试剂提供了试验依据。
Objective To investigate the specific allergen components of Dermatophagoides farinae. Methods ELISA assay was used to detect the bioactivity of allergen. The crude extract of Dermatoph- agoides farinae was prepared, and separated by SDS-PAGE to determine the relative molecular weight （MT） of every component. The allergic components were detected by Western blot with sera obtained from 60 dust mite allergic patients. Results There were 15 protein bands on the SDS-PAGE, the MT was from 14 × 10^3 to 109 × 10^3 , and 9 master tapes, the MT was 109 × 10^3, 100 × 10^3 , 86 × 10^3 , 62 × 10^3 , 56 × 10^3 , 36 × 10^3, 28 × 10^3, 19× 10^3 , 14 × 10^3 , respectively. Western blot assay showed that there were 5 bands of special allergen with the M, was 109 × 10^3, 100× 10^3, 36 × 10^3, 28 × 10^3, 14 × 10^3, respectively. The ELISA showed that the crude extract of Dermatophagoides farinae had bioactivity. Conclusion There are 5 special allergens in Dermatophagoidesfarinae with the MT of 109 × 10^3, 100 × 10^3, 36 × 10^3, 28 × 10^3 , 14× 10^3, respectively. Its extract has steady bioactivity. This study provides an evidence on the standardization of Derrnatophagoides farinae reagent for Chinese people.
出处
《中华微生物学和免疫学杂志》
CAS
CSCD
北大核心
2008年第8期690-692,共3页
Chinese Journal of Microbiology and Immunology
基金
广州市科技局重点攻关项目（2000-Z-025-01）
广东省科技厅重大专项（2005A10901003）
关键词
粉尘螨
变应原
免疫印迹
Dermatophagoides farinae
Allergen
Western blot
