1 Lynntech has developed a process to readily identify specific genomic loci in RNA-based bacteriophage and 2 viruses. We have demonstrated direct amplification of specific loci within the dengue genome using convective 3 RT-PCR (cRT-PCR). This direct detection of dengue eliminates the need for an independent sample 4 preparation step in the diagnosis of dengue infection. In doing so, we transitioned our convective amplification 5 units to perform RNA reverse transcription (RT), as well as DNA amplification (PCR). The convective assay is 6 based on the principle that two different temperatures at the ends of a cylinder will result in a buoyancy-driven 7 steady circulatory flow between those ends. Thus, PCR reagents in the cylinder will flow through a temperature 8 gradient allowing the necessary steps for amplification: denaturation, annealing, and elongation. Convective 9 amplification is rather appealing in that it requires very little power. Because reagents circulate within a 10 temperature gradient, there is no need for temperature ramping and power is not needed to cool, and then 11 heat, the reaction. So, our convective system can be powered by batteries and, therefore, provides a portable 12 means to detect and identify dengue infection, at the point-of-care. 13 Development of Lynntech?s dengue diagnostic includes the design and construction a disposable cartridge and 14 base unit, which allow the amplification of the dengue loci in point-of-care (and often, low resource) settings. 15 Dried reagents for the cRT-PCR reaction are contained within the cartridge, providing adequate reagent 16 stability under ambient storage conditions. The base unit is battery-operated, which makes it a perfect 17 diagnostic for field-use. Amplification products from the cRT-PCR reaction are applied to a lateral flow strip, 18 which is embedded in the cartridge, for analysis. This entire process takes less than one hour. Finally, it is 19 worth noting that Lynntech will meet with the FDA to discuss future studies that are necessary for regulatory 20 approval of our dengue diagnostic. 21 This application for an Administrative Supplement is to purchase equipment that will allow Lynntech to 22 manufacture lateral flow strips, in-house. This should represent significant savings with respect to further 23 development, testing, and commercialization of our dengue diagnostic. In addition, bringing lateral flow 24 development in-house will reduce the current risks associated with sole sourcing the technology. In this 25 Administrative Supplement application, we are requesting monies for equipment, supplies, and training.
Lynntech is developing a simple, sample-to-answer, point-of-care diagnostic for dengue virus, combining convective RT-PCR with lateral flow analysis. The assay will be capable of detecting and indentifying each of the four dengue serotypes in human samples. As a next generation tool for delivering diagnostic analyses into the hands of health-care professionals, our device will provide necessary resources where traditional tests are not available.
