Abstract：
Collecting and obtaining sufficient amount of airborne particles for multiple microbial component assessments can be challenging. A passive dust sampling device, the electrostatic dust fall collector (EDC) has been established for assessing airborne exposures including endotoxin and glucans. Recently, with advances in next-generation sequencing techniques, EDCs were used to collect microbial cells for DNA sequencing analysis to promote the study of airborne bacterial and fungal communities. However, low DNA yields have been problematic when employing passive sampling with EDC. To address this challenge, we attempted to increase the efficiency of extraction. We compared DNA extraction efficiency of bacterial components from EDCs captured on filters through filtration using five extraction techniques. By measuring the abundance, diversity and structure of bacterial communities using qPCR and amplicon sequencing targeting 16S rRNA genes, we found that two techniques outperformed the rest. Furthermore, we developed protocols to simultaneously extract both DNA and endotoxin from a single EDC cloth. Our technique promotes a high quality to price ratio and may be employed in large epidemiological studies addressing airborne bacterial exposure where a large number of samples is needed.
摘要：
收集和获得足够量的空气传播颗粒用于多种微生物组分评估可能是具有挑战性的。一种被动式粉尘采样装置，静电降尘收集器（EDC）已建立用于评估空气暴露，包括内毒素和葡聚糖。最近,随着下一代测序技术的进步，EDC用于收集微生物细胞进行DNA测序分析，以促进对空气传播的细菌和真菌群落的研究。然而,当使用EDC进行被动采样时，低DNA产量是有问题的。为了应对这一挑战，我们试图提高提取效率。我们使用五种提取技术比较了通过过滤在过滤器上捕获的EDC中细菌成分的DNA提取效率。通过测量丰度，使用qPCR和扩增子测序靶向16SrRNA基因的细菌群落的多样性和结构，我们发现两种技术胜过其余技术。此外，我们开发了从单个EDC布中同时提取DNA和内毒素的方案。我们的技术促进了高质量的价格比，可用于大型流行病学研究，以解决需要大量样品的空气细菌暴露问题。
