The long term objective is to develop methodologies and products to facilitate the commercialization of the polymerase chain reaction (PCR) in clinical diagnostics. It is crucial that sample preparation procedures prior to PCR amplification be simplified, and contamination of previously amplified nucleic acid sequences be minimized. In Phase I, a simple sample preparation method, based on the ability of serum transferrin to bind heme which otherwise inhibits PCR, has been developed for DNA targets in whole blood. This is currently being marketed as a sample preparation kit, AmpPrep(TM). The purpose of Phase II is to broaden the application of AmpPrep(TM) to include RNA targets in whole blood, and to formulate additional sample preparation products for other clinical specimens. The primary aims are: 1) Validate the AmpPrep(TM) method with actual clinical HIV-I, HTLV-I and HTLV-II blood samples. 2) Broaden the method for application to RNA-based PCR for the detection of HCV and bacteria in blood. 3) Modify the method for the detection of Lyme spirochete in CSF and urine samples. 4) Incorporate the isopsoralen mechanism for minimizing the problem of DNA carry-over, which was developed independently at HRI. The information gained from Phase II will be used to develop additional commercial sample preparation kits for the PCR.
