用 McAb及ELISA法测定人血清葎草花粉特异性IgE

目的 律草花粉是中国大部分地区夏秋季花粉症最重要的致敏花粉。本实验应用国产试剂和仪器建立测定血清律草花粉特异ＩｇＥ（ｓＩｇＥ）酶联有附试验方法。方法 采用ＥＬＩＳＡ方法,同时对３０例正常人和１４３例夏秋季花粉症患者进行检测。结果 患者组血清ｓＩｇＥ水平明显高于对照组;皮试反应程度与血清ｓＩｇＥ水平显著相关;皮试与血清ｓＩｇＥ的一致率为７４－８３％。经过敏原特异性抑制加热灭活试验本方法特异性和敏感怀

An ELISA assay for humulus scandens specific IgE antibodies

OBJECTIVE: Humulus scandens pollen (HSP) has been proved to be the one of the most important causative factors of autumnal hay fever in China. We established an enzyme immunoassay for specific IgE against HSP by using monoclonal anti-IgE antibody to detect sIgE in 143 patients with autumnal hay fever and 30 healthy volunteers. METHODS: An indirect method of ELISA was developed. The optimal conditions for the test were: coating the allergen at 4 degrees C overnight with a concentration of 12 micrograms/ml; the test serum was diluted 1:16 and incubated with allergen overnight at 37 degrees C; the most suitable dilution and incubative time of the peroxidase-conjugated mouse anti-human IgE antibody were 1:3,000 and 2 hours respectively. RESULTS: The patients' serum level of sIgE was significantly higher than controls (P < 0.001) and correlated significantly with the degree of skin test reaction (P < 0.01). The concordance between the results of skin test and ELISA was 74%-83%. The specificity of the assay was confirmed by heat inactivation and multiple absorption experiments. The coefficients of variation for the intraassay and interassay reproducibility ranged from 1.20%-8.30% and 8.63%-9.22% respectively. CONCLUSIONS: Determination of specific HSP IgE with ELISA assay shows good specificity, sensitivity and reproducibility. It favours clinical practice due to its lower cost.