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用于环境样品中(1→6)分支的、(1→3)-β-D-葡聚糖检测的酶联免疫吸附测定法。

Enzyme-linked immunosorbent assay specific for (1-->6) branched, (1-->3)-beta-D-glucan detection in environmental samples.

作者信息

Milton D K, Alwis K U, Fisette L, Muilenberg M

机构信息

Department of Environmental Health, Harvard School of Public Health, Boston, Massachusetts 02115, USA.

出版信息

Appl Environ Microbiol. 2001 Dec;67(12):5420-4. doi: 10.1128/AEM.67.12.5420-5424.2001.

Abstract

(1-->3)-beta-D-Glucans have been recognized as a potential causative agent responsible for bioaerosol-induced respiratory symptoms observed in both indoor and occupational environments. A specific enzyme immunoassay was developed to quantify (1-->6) branched, (1-->3)-beta-D-glucans in environmental samples. The assay was based on the use of a high-affinity receptor (galactosyl ceramide) specific for (1-->3)-beta-D-glucans as a capture reagent and a monoclonal antibody specific for fungal cell wall beta-D-glucans as a detector reagent. The assay was highly specific for (1-->6) branched, (1-->3)-beta-D-glucans (such as that from Saccharomyces cerevisiae) and did not show any response at 200 ng/ml to curdlan, laminarin, pustulan, dextran, mannan, carboxymethyl cellulose, and endotoxins. The detection level was 0.8 ng/ml for baker's yeast glucan and Betafectin. A coefficient of variation of 7.8% was obtained for (1-->3)-beta-D-glucans in house dust samples. Metal working fluids spiked with (1-->3)-beta-D-glucans inhibited the glucan assay. Because the assay is specific for (1-->6) branched, (1-->3)-beta-D-glucans and is sensitive and reproducible, it will be useful for the investigation of health effects from exposure to this class of biologically active molecules.

摘要

(1→3)-β-D-葡聚糖已被公认为是在室内和职业环境中观察到的生物气溶胶诱发呼吸道症状的潜在致病因素。开发了一种特定的酶免疫测定法来定量环境样品中的(1→6)分支的(1→3)-β-D-葡聚糖。该测定法基于使用对(1→3)-β-D-葡聚糖具有高亲和力的受体(半乳糖基神经酰胺)作为捕获试剂,以及对真菌细胞壁β-D-葡聚糖具有特异性的单克隆抗体作为检测试剂。该测定法对(1→6)分支的(1→3)-β-D-葡聚糖(如来自酿酒酵母的葡聚糖)具有高度特异性,在200 ng/ml时对短梗霉多糖、海带多糖、石耳多糖、葡聚糖、甘露聚糖、羧甲基纤维素和内毒素无任何反应。面包酵母葡聚糖和Betafectin的检测水平为0.8 ng/ml。室内灰尘样品中(1→3)-β-D-葡聚糖的变异系数为7.8%。添加了(1→3)-β-D-葡聚糖的金属加工液会抑制葡聚糖测定。由于该测定法对(1→6)分支的(1→3)-β-D-葡聚糖具有特异性,且灵敏且可重复,因此对于研究接触此类生物活性分子对健康的影响将很有用。

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