Pogner C-E, Gorfer M, Raulf M, Strauss J, Sander I
Bioresources, AIT Austrian Institute of Technology GmbH, Tulln, Austria.
Institute of the Ruhr-University Bochum (IPA), Institute for Prevention and Occupational Medicine of the German Social Accident Insurance, Bochum, Germany.
Appl Environ Microbiol. 2025 Aug 20;91(8):e0016325. doi: 10.1128/aem.00163-25. Epub 2025 Jul 16.
Airborne fungal spores are recognized for their human health impact, yet dose-response relationships remain undefined despite decades of bioaerosol sampling and analysis. In this article, we quantify immunologically active compounds, using six fungal-specific enzyme-linked immunosorbent assays (ELISAs) and compare them with cultivation, spore counting, and quantitative PCR (qPCR), using identical filter extract aliquots, various concentrations, single-organism aerosols, and mixes, in a controlled environment. Results showed high agreement between ELISAs and spore counting. By contrast, CFU enumeration and qPCR showed lower correlation with the other methods. Results of ELISAs showed high reproducibility of technical replicates and sampling duplicates, whereas CFU and qPCR results had high deviations. ELISA detection limits ranged from 10 to 100 spores/mL for larger spores (, ) and from 103 to 104 spores/mL for smaller spores (, , ). The and the ELISAs cross-reacted to or , respectively. The spore dust production method influenced the spore germination rate and qPCR results significantly. By contrast, ELISA results remain unchanged, with the exception of the ELISA. Quantitative spore counting was challenged by mixtures of species with morphologically similar spores from different fungal groups. In conclusion, the ELISA method is found suitable for fungal antigen quantification of air samples, provided the assays are sufficiently sensitive and specific, and germination evaluation is not required for risk assessment, leading to and ELISA recommendations.
Bioaerosol detection and analysis is an ongoing field of research. Although various methodologies are used for collection and analysis, there is no single method available to close the knowledge gap between exposure and health impact. For airborne fungal material, the standard analysis method remains cultivation. With molecular technology advancing in the field, both methods can only show the exposure to living, cultivatable, or total fungal cells in the airborne environment. To close the gap between airborne concentrations and impact on the human body, recognition of the allergenic potential is necessary. Therefore, we evaluated six fungal-specific ELISAs to make them ready for application in field studies and compared them to cultivation, spore counting, and molecular genetic methods. We are confident that in the future antigen-recognizing methods like the tested ELISAs will enable moving from particle detection toward detection of the disease-causing agent.
空气中的真菌孢子因其对人类健康的影响而受到关注，尽管经过数十年的生物气溶胶采样和分析，剂量反应关系仍不明确。在本文中，我们使用六种真菌特异性酶联免疫吸附测定（ELISA）对免疫活性化合物进行定量，并在受控环境中，使用相同的滤膜提取物等分试样、不同浓度、单一生物体气溶胶和混合物，将其与培养法、孢子计数法和定量聚合酶链反应（qPCR）进行比较。结果显示ELISA法与孢子计数法高度一致。相比之下，菌落形成单位（CFU）计数和qPCR与其他方法的相关性较低。ELISA结果显示技术重复和采样重复具有高度可重复性，而CFU和qPCR结果偏差较大。对于较大的孢子（ ， ），ELISA检测限为10至100个孢子/毫升，对于较小的孢子（ ， ， ），检测限为10³至10⁴个孢子/毫升。 ELISA和 ELISA分别与 或 发生交叉反应。孢子粉尘产生方法对孢子萌发率和qPCR结果有显著影响。相比之下，除 ELISA外，ELISA结果保持不变。定量孢子计数受到来自不同真菌组的形态相似孢子的物种混合物的挑战。总之，发现ELISA方法适用于空气样本的真菌抗原定量，前提是该检测足够灵敏和特异，且风险评估不需要萌发评估，从而得出 ELISA和 ELISA的建议。
生物气溶胶检测与分析是一个不断发展的研究领域。尽管使用了各种方法进行收集和分析，但尚无单一方法能够弥合暴露与健康影响之间的知识差距。对于空气中的真菌物质，标准分析方法仍然是培养法。随着该领域分子技术的进步，这两种方法都只能显示在空气环境中对活的、可培养的或总真菌细胞的暴露情况。为了弥合空气中的浓度与对人体的影响之间的差距，识别致敏潜力是必要的。因此，我们评估了六种真菌特异性ELISA，使其准备好在现场研究中应用，并将它们与培养法、孢子计数法和分子遗传学方法进行比较。我们相信，未来像经过测试的ELISA这样的抗原识别方法将能够从颗粒检测转向致病因子的检测。
