Portable platform for rapid in-field identification of human fecal pollution in water.
Portable platform for rapid in-field identification of human fecal pollution in water.
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DOI：
10.1016/j.watres.2017.12.023
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发表时间：
2018-03-15
期刊：
影响因子：
12.8
通讯作者：
Bhadra S
中科院分区：
文献类型：
--
作者：
Jiang YS;Riedel TE;Popoola JA;Morrow BR;Cai S;Ellington AD;Bhadra S
人类粪便对水的污染是一种公共卫生风险。然而，不充分的测试解决方案阻碍了及时的、可操作的监控。基于细菌培养的方法简单，但通常无法区分粪便宿主来源。PCR检测可以识别宿主来源，但需要专业知识和基础设施。为了弥补这一差距，我们开发了一种现场可用的核酸诊断平台和快速样品制备方法，可以在采样后80分钟内现场识别人类粪便污染。我们的平台依赖于来自粗样品的人相关拟杆菌HF 183遗传标记的环介导等温扩增（LAMP）。寡核苷酸链交换（OSD）探针通过序列特异性地将LAMP扩增子转导成可由未修饰的智能手机拍摄的可见荧光来减少假阳性。我们的检测方法可以在污水污染的水中检测到低至17拷贝/ml的人类相关HF 183靶点，而不会与犬或猫的粪便发生交叉反应。它对各种环境水源和未经处理的污水都有很好的处理效果。我们还开发了冻干分析和廉价的3D打印设备，以最大限度地降低成本并促进现场应用。
Human fecal contamination of water is a public health risk. However, inadequate testing solutions frustrate timely, actionable monitoring. Bacterial culture-based methods are simple but typically cannot distinguish fecal host source. PCR assays can identify host sources but require expertise and infrastructure. To bridge this gap we have developed a field-ready nucleic acid diagnostic platform and rapid sample preparation methods that enable on-site identification of human fecal contamination within 80 min of sampling. Our platform relies on loop-mediated isothermal amplification (LAMP) of human-associated Bacteroides HF183 genetic markers from crude samples. Oligonucleotide strand exchange (OSD) probes reduce false positives by sequence specifically transducing LAMP amplicons into visible fluorescence that can be photographed by unmodified smartphones. Our assay can detect as few as 17 copies/ml of human-associated HF183 targets in sewage-contaminated water without cross-reaction with canine or feline feces. It performs robustly with a variety of environmental water sources and with raw sewage. We have also developed lyophilized assays and inexpensive 3D-printed devices to minimize cost and facilitate field application.
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影响因子：
2.7
作者：
Abbasi I;Kirstein OD;Hailu A;Warburg A
通讯作者：
Warburg A
影响因子：
3.4
作者：
Haugland, Richard A.;Varma, Manju;Shanks, Orin C.
通讯作者：
Shanks, Orin C.
DOI：
10.1099/00221287-137-7-1565
发表时间：
1991-07-01
期刊：
JOURNAL OF GENERAL MICROBIOLOGY
影响因子：
--
作者：
CAMPBELL, S;YATES, PS;RICHMOND, SJ
通讯作者：
RICHMOND, SJ
影响因子：
3.4
作者：
Gasalla, Maria A.;Gandini, Fabricio C.
通讯作者：
Gandini, Fabricio C.
影响因子：
5.4
作者：
Crocker, Jonny;Shields, Katherine F.;Venkataramanan, Vidya;Saywell, Darren;Bartram, Jamie
通讯作者：
Bartram, Jamie