Institute for Prevention and Occupational Medicine of the German Social Accident Insurance, Institute of the Ruhr University Bochum (IPA), Bochum, Germany.
J Toxicol Environ Health A. 2012;75(19-20):1185-93. doi: 10.1080/15287394.2012.707603.
The aim of our study was to develop specific enzyme-linked immunosorbent assays (ELISAs) and apply these to assess mold antigen exposure in composting plants. Sandwich ELISAs based on polyclonal antibodies to Aspergillus fumigatus (Af), Penicillium chrysogenum (Pc), and Cladosporium herbarum (Ch) antigens were developed and validated. Reactivity to 18 different mold species was tested. To optimize extraction procedure, inhalable dust samples taken by a parallel sampler were extracted with or without homogenization. In 31 composting plants stationary pumps were installed at 4 sites to collect 124 inhalable dust samples. The newly developed ELISAs were used in addition to an anti β-1,3-glucan ELISA to quantify mold antigens. The Cladosporium ELISA showed less than 0.04% reactivity to extracts from other fungal genera, while the Af ELISA demonstrated a reactivity of up to 3.6% and the Pc ELISA reacted up to 11% to other mold species. Extraction of parallel sampled filters gave higher antigen amounts with homogenization. The increase was highest for Pc-antigens, followed by Af-antigens, and lowest for Ch-antigens. Mean lower detection limits of homogenized inhalable dust samples were 5 ng/m(3) (Af), 0.6 ng/m(3) (Pc), 0.2 ng/m(3) (Ch), and 0.6 ng/m(3) (β-1,3-glucan). The ELISAs were able to detect antigens in 43% (Af), 37% (Pc), 94% (Ch), or 100% (β-1,3-glucan) of the 124 airborne dust samples. Inhalable dust, β-1,3-glucan, and Af-, Pc-, and Ch-antigen concentrations were significantly correlated. The newly developed mold antigen ELISAs are thus able to measure airborne exposure levels in composting plants and differentiate between distinct fungi genera.
本研究旨在开发特定的酶联免疫吸附测定法（ELISA），并应用这些方法来评估堆肥厂中霉菌抗原的暴露情况。我们开发并验证了基于多克隆抗体对烟曲霉（Af）、产黄青霉（Pc）和枝孢菌（Ch）抗原的夹心 ELISA。测试了对 18 种不同霉菌的反应性。为了优化提取程序，使用平行采样器采集的可吸入粉尘样品进行了或不进行匀浆提取。在 31 个堆肥厂中，在 4 个地点安装了固定泵，以收集 124 个可吸入粉尘样品。除了抗β-1,3-葡聚糖 ELISA 之外，我们还使用新开发的 ELISA 来定量霉菌抗原。枝孢菌 ELISA 对其他真菌属的提取物的反应性小于 0.04%，而 Af ELISA 的反应性高达 3.6%，Pc ELISA 的反应性高达 11%至其他霉菌。用匀浆提取平行采样器的滤器可获得更高的抗原量。增加量最高的是 Pc 抗原，其次是 Af 抗原，最低的是 Ch 抗原。匀浆可吸入粉尘样品的平均下限检测值分别为 5 ng/m³（Af）、0.6 ng/m³（Pc）、0.2 ng/m³（Ch）和 0.6 ng/m³（β-1,3-葡聚糖）。ELISA 能够检测到 124 个空气中粉尘样本的 43%（Af）、37%（Pc）、94%（Ch）或 100%（β-1,3-葡聚糖）中存在抗原。可吸入粉尘、β-1,3-葡聚糖以及 Af、Pc 和 Ch 抗原浓度呈显著相关。因此，新开发的霉菌抗原 ELISA 能够测量堆肥厂中的空气暴露水平，并区分不同的真菌属。
