Inexpensive, Rapid and Comprehensive Virulence and Marker Gene (VMG) Analyzer for Waterborne Pathogens

Every year, an estimated 19.5 million illnesses occur in the United States due to consumption of unsafe drinking water and result in productivity losses of the order of $20 billion. The burden of waterborne illnesses is expected to increase due to the emergence of more virulent microorganisms and an increase in the number of sensitive subpopulations, which include older people, young children, pregnant women, and immunocompromised persons. Conventional culturing of indicator organisms is known to be inadequate for assessing the presence and concentration of specific waterborne pathogens. Newer molecular assays involving real-time PCR and microarrays allow sensitive and specific detection of multiple pathogens simultaneously, but require expensive and bulky PCR thermocyclers and microarray fluorescence scanners, and involve many manual procedures. Consequently, these tests are only available in well-equipped laboratories with highly trained personnel. Hence, more affordable, rapid, rugged, and portable devices for microbiological water quality monitoring are critically needed. To address this need, AuqaBioChip LLC (Lansing, MI) is focused on bringing to the market an inexpensive, easy-to-use, and portable device (referred to as the VMG Analyzer) for rapid molecular detection of waterborne pathogens. This device will be able to analyze water samples for multiple pathogens simultaneously at low cost by integrating loop-mediated amplification (LAMP) and microarray hybridization in disposal of polymer microfluidic chips. It will be provided at a cost that is substantially lower than that of current equipment required for molecular detection of waterborne pathogens, and the cost per assay will be less than $5 due to the use of polymer microfluidic chips and the more than 10-fold reduction in reagent volume compared to conventional assays. The assay will provide results with high analytical accuracy by coupling LAMP-based amplification of multiple target genes and microarray-based analysis of the amplicons in approximately 30 minutes. In this Phase I project, AquaBioChip LLC plans to develop and evaluate the LAMP-microarray microfluidic chips for detection of eight waterborne pathogens that are linked to the majority of waterborne disease outbreaks in the United States.