One-Step, POC Sample-to-Answer Process for RNA Analysis Outside the Laboratory

PROJECT SUMMARYLynntech has developed a process to readily identify specific genomic loci in RNA based bacteriophage andvirusesWe have demonstrated direct amplification of specific loci within the dengue genome using convectiveRT PCRcRT PCRThis direct detection of dengue eliminates the need for an independent samplepreparation step in the diagnosis of dengue infectionIn doing sowe transitioned our convective amplificationunits to perform RNA reverse transcriptionRTas well as DNA amplificationPCRThe convective assay isbased on the principle that two different temperatures at the ends of a cylinder will result in a buoyancy drivensteady circulatory flow between those endsThusPCR reagents in the cylinder will flow through a temperaturegradient allowing the necessary steps for amplificationdenaturationannealingand elongationConvectiveamplification is rather appealing in that it requires very little powerBecause reagents circulate within atemperature gradientthere is no need for temperature ramping and power is not needed to cooland thenheatthe reactionSoour convective system can be powered by batteries andthereforeprovides a portablemeans to detect and identify dengue infectionat the point of careDevelopment of Lynntech s dengue diagnostic includes the design and construction a disposable cartridge andbase unitwhich allow the amplification of the dengue loci in point of careand oftenlow resourcesettingsDried reagents for the cRT PCR reaction are contained within the cartridgeproviding adequate reagentstability under ambient storage conditionsThe base unit is battery operatedwhich makes it a perfectdiagnostic for field useAmplification products from the cRT PCR reaction are applied to a lateral flow stripwhich is embedded in the cartridgefor analysisThis entire process takes less than one hourFinallyit isworth noting that Lynntech will meet with the FDA to discuss future studies that are necessary for regulatoryapproval of our dengue diagnosticThis application for an Administrative Supplement is to purchase equipment that will allow Lynntech tomanufacture lateral flow stripsin houseThis should represent significant savings with respect to furtherdevelopmenttestingand commercialization of our dengue diagnosticIn additionbringing lateral flowdevelopment in house will reduce the current risks associated with sole sourcing the technologyIn thisAdministrative Supplement applicationwe are requesting monies for equipmentsuppliesand training Lynntech is developing a simplesample to answerpoint of care diagnostic for dengue viruscombining convective RT PCR with lateral flow analysisThe assay will be capable of detecting and indentifying each of the four dengue serotypes in human samplesAs a next generation tool for delivering diagnostic analyses into the hands of health care professionalsour device will provide necessary resources where traditional tests are not available