摘要
将6只美洲大蠊全虫用液氮研磨提取粗浸液,用阴离子交换剂(DEAE,SephadexA-50)纯化后测定蛋白浓度,并配制成不同浓度梯度,分别将粗浸液及纯化后的不同浓度变应原点于硝酸纤维膜(NC膜)上,依次加入不同蟑螂过敏患者血清、生物素-亲和素系统的IgE二抗和辣根过氧化物酶(HRP),以及鲁米诺化学发光底物进行化学发光反应。结果表明,化学发光免疫法可检测美洲大蠊粗浸液最低蛋白浓度为0.87μg/ml。在此浓度下,其检出结果与皮试阳性患者血清符合率为90%,与皮试阴性及健康人血清的符合率均为100%。
Crude extract of Periplaneta americana was prepared by liquid nitrogen grinding. After being purified with DEAE Sephadex A-50 ion exchange chromatography, the protein content of the extract was determined and the extract solution was prepared at gradient concentrations. The crude extract and purified allergen at different concentrations were dotted respectively on nitrocellulose （NC） membrane. Patient serum, bio-IgE, sa-HRP, luminal regents were added to the membrane. The chemiluminescence was displayed by exposing to X-film. The result revealed that the minimum protein content of crude Periplaneta americana extract detected by CLIA is 0.87μg/ml, with 90% accordance to skin test positive patients, and 100% accordance to those with negative skin test and ELISA detection.
出处
《中国寄生虫学与寄生虫病杂志》
CAS
CSCD
北大核心
2006年第6期471-473,共3页
Chinese Journal of Parasitology and Parasitic Diseases
基金
国家自然科学基金(No.39860071
30571625)
广东省科技计划重点项目(No.2003A3080502)
深圳市科技计划项目(No.200326)~~
