A safer framework to evaluate characterization technologies of exhaled biologic materials using electrospun nanofibers.
A safer framework to evaluate characterization technologies of exhaled biologic materials using electrospun nanofibers.
复制标题
使用电纺纳米纤维评估呼出生物材料表征技术的更安全框架。
DOI：
10.1039/d3nr01859h
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发表时间：
2023
期刊：
影响因子：
6.7
通讯作者：
Haselton,FrederickR
中科院分区：
文献类型：
--
作者：
Evans,DavidT;Nelson,DaltonJ;Pask,MeganE;Haselton,FrederickR
呼出的生物物质是许多呼吸道感染传播的来源。为了避免管理危险病原体所需的高水平生物安全性，我们开发了一个更安全的框架，使用内源性替代目标 RNase P 和轻链球菌作为呼出生物制剂的采样手段。我们的呼气收集方案使用纳米级纤维状聚（乙烯醇）基材作为面罩插入物。经过一段时间的呼吸或说话后，插入物就会被移除并溶解。提取 RNase P RNA 和 S. mitis DNA，通过多重 RT-qPCR 进行定量。在呼吸至少五分钟或说话一分钟期间获得的所有样本中都检测到了这两种替代生物标志物。重复 30 次的短语具有最多的拷贝，其中有 375 ± 247 个 S. mitis 和 54 ± 33 个 RNase P。当这些短语仅重复 5 次时，收集到的 S. mitis 拷贝仍然可检测到，但水平明显较低，S. mitis 为 11 ± 5，RNase P 为 12 ± 9。这些结果证明了一个收集和量化框架，可以很容易地适应进一步表征呼气来自健康个体的纳米级生物材料，安全地探索新的收集设计，并作为一种将样本控制纳入未来病原体呼出研究的方法。
Exhaled biologic material is the source for the spread of many respiratory tract infections. To avoid the high-level of biosafety required to manage dangerous pathogens, we developed a safer framework using the endogenous surrogate targets RNase P and Streptococcus mitis as a means to sample exhaled biologics. Our exhalation collection scheme uses nanoscale fibrous poly(vinyl alcohol) substrates as facemask inserts. After a period of breathing or speaking, the inserts are removed and dissolved. RNase P RNA and S. mitis DNA are extracted for quantification by multiplexed RT-qPCR. Both surrogate biomarkers were detected in all samples obtained during breathing for at least five minutes or speaking for one minute. Phrases repeated 30 times had the most copies with 375 ± 247 of S. mitis and 54 ± 33 of RNase P. When the phrases were repeated just 5 times, the S. mitis copies collected were still detectable but at a significantly lower level of 11 ± 5 for S. mitis and 12 ± 9 for RNase P. These results demonstrate a collection and quantification framework that can be readily adapted to further characterize the exhalation of nanoscale biologic materials from healthy individuals, explore new collection designs safely, and serve as a method to incorporate sample controls for future pathogen exhalation studies.