Particle Diffusometry: An Optical Detection Method for Vibrio cholerae Presence in Environmental Water Samples
Autor: | Dong Hoon Lee, Jacqueline C. Linnes, Katherine N. Clayton, Taylor J. Moehling, Tamara L. Kinzer-Ursem, Steven T. Wereley |
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Jazyk: | angličtina |
Rok vydání: | 2019 |
Předmět: |
0301 basic medicine
Microbiological Techniques lcsh:Medicine Biosensing Techniques medicine.disease_cause Polymerase Chain Reaction Fluorescence Article Diffusion 03 medical and health sciences 0302 clinical medicine medicine Sample preparation Particle Size lcsh:Science Vibrio cholerae Detection limit Multidisciplinary Chromatography Chemistry Viscosity lcsh:R 030104 developmental biology Nucleic acid Particle lcsh:Q Particle size Water Microbiology Biosensor Nucleic Acid Amplification Techniques 030217 neurology & neurosurgery |
Zdroj: | Scientific Reports, Vol 9, Iss 1, Pp 1-12 (2019) Scientific Reports |
ISSN: | 2045-2322 |
Popis: | There is a need for a rapid, robust, and sensitive biosensor to identify low concentrations of pathogens in their native sample matrix without enrichment or purification. Nucleic acid-based detection methods are widely accepted as the gold standard in diagnostics, but robust detection of low concentrations of pathogens remains challenging. Amplified nucleic acids produce more viscous solutions, which can be measured by combining these products with fluorescent particles and measuring the change in the particle diffusion coefficient using a technique known as particle diffusometry. Here, we utilize Vibrio cholerae (V. cholerae) as a proof-of-concept for our detection system due to its inherently low concentration in environmental water samples. We demonstrate that particle diffusometry can be used to detect down to 1 V. cholerae cell in molecular-grade water in 20 minutes and 10 V. cholerae cells in pond water in just 35 minutes in 25 µL reaction volumes. The detection limit in pond water is environmentally relevant and does not require any enrichment or sample preparation steps. Particle diffusometry is 10-fold more sensitive than current gold standard fluorescence detection of nucleic acid amplification. Therefore, this novel measurement technique is a promising approach to detect low levels of pathogens in their native environments. |
Databáze: | OpenAIRE |
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