Abstract
We report a rapid "sample-to-answer" platform that can be used for the quantitative monitoring of genetic biomarkers within communities through the analysis of wastewater. The assay is based on the loop-mediated isothermal amplification (LAMP) of nucleic acid biomarkers and shows for the first time the ability to rapidly quantify human-specific mitochondrial DNA (mtDNA) from raw untreated wastewater samples. mtDNA provides a model population biomarker associated with carcinogenesis including breast, renal and gastric cancers. To enable a sample-to-answer, field-based technology, we integrated a filter to remove solid impurities and perform DNA extraction and enrichment into a low cost lateral flow-based test. We demonstrated mtDNA detection over seven consecutive days, achieving a limit of detection of 40 copies of human genomic DNA per reaction volume. The assay can be performed at the site of sample collection, with minimal user intervention, yielding results within 45 min and providing a method to monitor public health from wastewater.
Highlights
Wastewater-based epidemiology has recently been shown to be an innovative and promising tool for the estimation of community-wide drug,[1,2] alcohol,[3] and tobacco use,[4] as well as for the monitoring of oxidative stress biomarkers such as F2-isoprostanes.[5]
We demonstrate a portable loop-mediated isothermal amplification (LAMP) assay to detect human-specific mitochondrial DNA (mtDNA) in wastewater, as a population biomarker, with the potential to monitor public health
We designed and demonstrated a LAMP assay for the detection of human specific mtDNA in wastewater, a potential population biomarker associated with cancer
Summary
Wastewater-based epidemiology has recently been shown to be an innovative and promising tool for the estimation of community-wide drug,[1,2] alcohol,[3] and tobacco use,[4] as well as for the monitoring of oxidative stress biomarkers such as F2-isoprostanes.[5]. In order to estimate the public health status using these methods, wastewater-based epidemiology requires human and disease-specific biomarkers. The criteria for such candidate biomarkers have been outlined previously.[8] Briefly, they should be excreted in bodily fluids predictably, as well as being detectable and stable in wastewater. Such biomarker proxies must be unique to humans, and of low variance in the per capita rate of daily excretion. The change of DNA sequence and structure via processes, such as damage, repair, and mutation, might provide more subtle indications of the evolution of disease at population level.[8,12]
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