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一种新型免疫分析法,可定量检测室内霉菌 Aspergillus versicolor 的真菌抗原。

A new immunoassay to quantify fungal antigens from the indoor mould Aspergillus versicolor.

机构信息

Institute for Prevention and Occupational Medicine of the German Social Accident Insurance, Institute of the Ruhr-Universität Bochum IPA, Center of Allergology/Immunology, Buerkle-de-la-Camp-Platz 1, 44789 Bochum, Germany.

出版信息

Environ Sci Process Impacts. 2013 Jun;15(6):1162-71. doi: 10.1039/c3em30870g.

Abstract

Aspergillus versicolor is among the most commonly found moulds in moisture-damaged buildings and can be associated with adverse health effects in humans. This paper reports the development, validation and application of an enzyme immunoassay to quantify A. versicolor antigens. A sandwich ELISA was developed using polyclonal antibodies that recognize a broad range of A. versicolor proteins present in fungal spores and in mycelia fragments. To validate the new method, A. versicolor antigens were quantified in samples collected from homes with visible mould growth, including dust from vacuumed walls and bulk samples of building materials. Antigen concentrations were compared to the results of a commercial ELISA based on monoclonal antibodies (AveX ELISA, Indoor Biotechnologies, Charlottesville, USA) and correlated with colony forming units (CFU) of A. versicolor. The A. versicolor ELISA was very sensitive with a lower detection limit of 120 pg ml(-1). The assay also showed some reactivity to other moulds with strongest reactions with other Aspergillus species (1-3% reactivity). The new assay detected A. versicolor antigens in a much higher percentage of dust samples (88% vs. 27%) and bulk samples (89% vs. 24%) than the AveX assay. A significant correlation (r = 0.67, and p < 0.0001) was found between antigen concentrations and CFU of A. versicolor. Based on its low detection limit and good correlation with the culture-based method, this new immunoassay seems to be a useful tool for the measurement of A. versicolor exposure levels and a reliable complement to the traditional monitoring techniques, such as mould cultivation or microscopy.

摘要

杂色曲霉菌是在潮湿受损建筑物中最常见的霉菌之一,可与人类的不良健康影响有关。本文报告了开发、验证和应用一种酶联免疫吸附测定法来定量杂色曲霉菌抗原的方法。使用针对真菌孢子和菌丝体片段中存在的广泛的杂色曲霉菌蛋白的多克隆抗体开发了夹心 ELISA。为了验证新方法,在有可见霉菌生长的家庭中收集的样本中定量了杂色曲霉菌抗原,包括从真空墙壁吸尘的灰尘和建筑材料的大块样本。将抗原浓度与基于单克隆抗体的商业 ELISA(AveX ELISA,美国夏洛茨维尔室内生物技术公司)的结果进行比较,并与杂色曲霉菌的菌落形成单位(CFU)相关联。杂色曲霉菌 ELISA 非常敏感,检测下限为 120 pg ml(-1)。该测定法还显示出对其他霉菌的一些反应性,与其他曲霉属物种的反应最强(1-3%的反应性)。新测定法比 AveX 测定法检测到更高比例的灰尘样本(88%比 27%)和大块样本(89%比 24%)中的杂色曲霉菌抗原。抗原浓度与杂色曲霉菌 CFU 之间存在显著相关性(r = 0.67,p < 0.0001)。基于其低检测限和与基于培养的方法的良好相关性,这种新的免疫测定法似乎是测量杂色曲霉菌暴露水平的有用工具,也是对传统监测技术(如霉菌培养或显微镜检查)的可靠补充。

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