摘要
目的 应用Western blotting的方法检测蚕蛹（silkworm chrysalis）浸出液中致敏原的成份，为蚕蛹致敏原的研究和临床诊断提供依据。方法取蚕蛹浸出液，经SDS-聚丙烯酰胺凝胶分离，再转印到PVDF膜上，封闭后将血清与膜条共同孵育，后与辣根过氧化物酶标记的羊抗人IgE抗体反应，显色底物用对氨基联苯胺。结果SDS-PAGE显示蚕蛹可辨条带有12条，分子量在14～94kD之间，其中主带有8条，分子量依次为80kD、68kD、62kD、50kD、29kD、23KD、18kD、16kD。Western blotting结果表明，22例蚕蛹过敏患者血清全部呈阳性反应，浸出液中共有6条致敏条带，其中分子量68kD和29kD是主要致敏组份，阳性反应率均为100％。结论蚕蛹分子量68KD和29kD的组份为主要致敏组份，结果可为开发适舍我国特色的蚕蛹变应原提供依据。
Objective To analyze silkworm chrysalis allergens using the method d Western blotting and provide the information for further research and clinical diagnosis.Methods The crude extract were separated on SDS-PAGE and transferred to PVDF membrane.After blocking,the membrane strips were incubated with patients＇ sera and HRP conjugated goat anti-human IgE antibody respectively.The bands were detected by DAB Results SDS-PAGE analysis revealed that the proteins of silkworm chrysalis comprised of at least twelve discrete protein bands with the molecular weight ranging from 14 kD to 94 kD, and the molecular weight d each band is 80 kD, 68 kD,62.1 kD,50 kD, 29 kD, 23 kD, 18 kD, 16 kD respectively. Western blot assay showed that six allergen bands separated from the crude extract were reacting with sere obtained from 22 crab ailergenic subjects,and the bards d 68 and 29 kD were the major allergenic compents.The positive rate was 100%. Conclusion The 68 kD and 29 kD bands are the major allergens of silkworm chrysalis. The study has laid a base for the further purifcation and standardization of silkworm chrysalis allergen suitable to Chinese people.
出处
《广西医学》
CAS
2006年第1期43-45,共3页
Guangxi Medical Journal
