Materials for Isolation of Nucleic Acids from Whole Blood without a Lysis Step

DESCRIPTION (provided by applicant): The objective of the proposed study is to develop new materials for the rapid isolation of nucleic acids from complex matrices such as whole blood. In Phase I we will prepare functionalized \microparticles for binding and releasing nucleic acids that utilize proprietary chemistry recently i developed at Lumigen. These materials find many uses but are intended particularly for the rapid ' and simplified isolation of genomic nucleic acids from unpurified or heterogeneous matrices, especially DNA in mammalian tissue or whole blood. This is made possible by a unique feature of these materials: their ability to capture DNA and/or RNA under virtually any reasonable conditions. As a result we have been able to bind DNA from whole blood without any lysis step. The prototype new materials bind nucleic acids with exceptional strength under a range of conditions and resist release under a wide variety of conditions. The unique binding properties of the new materials will be utilized to develop optimized methods for stringently washing and releasing the bound nucleic acid in a highly controllable fashion. Preliminary experiments have demonstrated that a cleavable link between the solid support and the binding moiety can be controllably severed to release nucleic acid back into solution. Nucleic acids purified with first generation materials have been found compatible with downstream applications including amplification technologies such as PCR and LMO, a ligase-based nucleic acid amplification technology invented by the PL We expect to produce one or more general use kits for isolating DNA from whole blood without any lysis step and in a quantity and quality acceptable for use in downstream amplification and diagnostic applications. For the initial phase of the project we expect to develop both non-magnetic and | magnetically responsive microparticles from different solid phase support materials. Since the j prototype new materials have shown exceptionally strong binding to nucleic acids, we plan further to begin exploring the use of the developed materials in capturing nucleic acids from low abundance viral and bacterial infectious agents from samples such as blood and other body fluids. This work serves as a prelude to the development of nucleic acid-based test methods for pathogens in human blood and food products to be undertaken in Phase II.