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高通量定量PCR的验证和应用,用于在环境水中同时检测微生物源跟踪标记物

Sunayana Raya¹, Bikash Malla², Ocean Thakali¹

¹Department of Engineering, University of Yamanashi, 4-3-11 Takeda, Kofu, Yamanashi 400-8511, Japan.

The Science of the total environment

|May 31, 2024

概括

这项研究验证了微生物源追踪 (MST) 的高通量定量聚合酶链反应 (HT-qPCR). 该方法成功检测到水中的多种便污染源,细菌标记物显示出高灵敏度和精度.

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:

微生物源追踪 (MST) 对于识别水体中的便污染源至关重要.
没有一个MST标记在所有水类型中普遍适用.
开发用于同时检测多个MST标记物的方法至关重要.

研究的目的:

验证一种高通量定量聚合酶连锁反应 (HT-qPCR) 方法,用于同时检测多个MST标记物.
使用HT-qPCR评估各种宿主特异性MST标记物的准确性和适用性.
在尼泊尔加德满都山谷的各种水体中识别便污染源.

主要方法:

使用HT-qPCR与26个便来源样本验证10个宿主特异性MST标记物 (细菌体,线粒体DNA,病毒).
应用九个经过验证的标记物来分析地下水,油轮加油站,饮用水处理厂和河水中的便污染.
将HT-qPCR结果与标准定量聚合酶连锁反应 (qPCR) 的比较.

主要成果:

狗线粒体DNA (mtDNA) 标记实现了100%的准确性;所有细菌标记显示了100%的灵敏度.
在河水样本中,人类特异性细菌菌 (BacHum) 和反动物特异性细菌菌 (BacR) 标志物非常普遍 (分别为83%和100%).
HT-qPCR结果与标准qPCR有很强的一致性,证实了该方法的可靠性.

结论:

HT-qPCR是一种可靠和有效的方法,用于同时检测多个微生物源跟踪标记物.
经过验证的MST标记物显示出在各种水类中识别各种便污染源的潜力.
这种方法增强了加德满都谷和类似地区便污染的水质监测和管理策略.

关键词:

饮用水来源饮用水来源便污染便污染高通量 qPCRR 的使用.微生物源追踪微生物源追踪尼泊尔尼泊尔尼泊尔尼泊尔尼泊尔

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