This application proposes the use of a methodology, immobilized sample amplification (ISA), to achieve PCR amplification of nucleic acids in clinical samples, eliminating the need for a nucleic acid purification step. Preliminary data are presented that indicate that PCR amplification of unfixed, immobilized blood cells can be achieved. The goal of this Phase I application is to demonstrate that ISA is a sensitive and reproducible method to detect viral RNA in blood, using the equine infectious anemia virus (EIAV) as a paradigm. For this purpose, the cellular and plasma portion of blood will be immobilized on a filtration membrane. The method will utilize a proprietary disposable device, the Gene Flo chamber, to collect a very small amount of blood and dry it into a reaction chamber where amplification and detection of the target sequences takes place.
Thesaurus Terms:blood test, communicable disease diagnosis, diagnosis design /evaluation, polymerase chain reaction equine infectious anemia virus, human immunodeficiency virus, virus RNA human tissueNational Heart, Lung and Blood Institute (NHLBI)
