胺和基于总DNA的qPCR和长读序列测序的比较,用于在河口河流中追踪微生物来源
Genevieve Ehasz1, Lylla Almosd1, Piash Ahamed1
1Civil & Environmental Engineering, Rutgers University, 500 Bartholomew Rd., Piscataway, NJ 08854, United States of America.
The Science of the total environment
|December 28, 2025
概括
一氧化 (PMA) -DNA方法显示,通过与可培养微生物比总DNA更好地相关联,更准确的源跟踪具有前途. 这项研究比较了PMA-DNA和总DNA,用于跟踪下拉里丹河中人类便污染.
科学领域:
- 环境微生物学环境微生物学
- 评估水质水质的评估.
- 分子生物学技术分子生物学技术.
背景情况:
- 病原体污染是一个主要的水质问题,推动了有效的便源追踪 (FST) 的需求,以指导缓解努力.
- 目前的生物分子FST方法,如qPCR和基于图书馆的方法,由于水生环境中持续存在的DNA,可以产生错误的阳性结果.
- 使用可活细胞DNA (propidium monoazide,PMA-DNA) 来改善与可培养便微生物的相关性,比总DNA方法更少探索.
研究的目的:
- 为了比较总DNA与PMA-DNA在人类便标记物HF183 qPCR和长期读取的基于图书馆的FST中的有效性.
- 评估生物分子FST结果与公民科学监测中的肠球菌观察结果之间的相关性.
- 评估不同DNA制备方法对FST准确性和来源归因的影响.
主要方法:
- 对比人类便标志物HF183 qPCR和长读纳米孔测序用于FST,使用总DNA和PMA-DNA.
- 收集了新泽西州下拉里坦河 (Lower Raritan River, NJ) 的非游泳海的水样,用于肠球菌分析.
- 使用 SourceTracker 软件,根据基于库的 FST 数据进行源代码归因.
主要成果:
- 与PMA-DNA相比,与总DNA相比,观察到较低的HF183基因拷贝度,表明总DNA对人类便贡献的潜在高估.
- 对于PMA-DNA或总DNA,没有发现HF183和肠球菌之间的相关性.
- 与总DNA相比,PMA-DNA在假定便来源和肠球菌观察之间产生了更强的相关性.
- 来源归因因取样地点和DNA制备方法而异;总离心机缩DNA与肠球相关,与总过器缩DNA不同.
结论:
- PMA-qPCR和PMA-纳米孔测序对FST具有实用性,可能为可行的便污染提供更准确的评估.
- 选择DNA制备方法显著影响FST结果和来源归因准确度.
- 需要进一步的研究来评估PMA-DNA方法的成本效益和劳动力要求,以进行常规的FST.
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