Mr Meysam Khodaparast1
1AgriBio, La Trobe University, Melbourne/Bundoora, Australia
Supplying healthy water is significant in protecting human health as the growing population continues to deteriorate water quality. One of the main elements of water quality degradation is human faecal pollution via spreading pathogens in waterways. Therefore, quick diagnosis of human faecal contamination in environmental water is a priority in protecting public health from waterborne-pathogens outbreaks worldwide. The current methods of detecting pathogens, such as qPCR and culturing techniques, are timely and complicated. The requirement of completing these assays in a centralised laboratory with expertise creates time and cost challenges.
In this study, we optimised a Loop-mediated isothermal amplification assay (LAMP) to detect Bacteroides DNA as an indicator of human faecal contamination in environmental water. The most challenging step in eDNA analysis- eDNA sampling and purification- was optimised through a water sample filtration combined with a lysis buffer enabling to perform the assay in-field. The Bac-LAMP assay developed in this study can reliably detect 2 CFU μL−1 in a time to positive of under 10 minutes. This highly sensitive and specific assay was tested on Victoria’s waterways and demonstrated the potential to be substituted with the current qPCR to detect pathogens in environmental waters. With the simplified sample preparation, the rapid and reliable Bac-LAMP assay can assist water regulators in quickly assessing water quality on the site to protect public health.
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