Abstract: Disclosed is a novel method for diagnosis or differential diagnosis of disease agents and secondary disease agents. The method disclosed uses a novel amplification strategy termed TemPCR to allow sensitive and specific amplification of target sequences from any disease agents and/or secondary disease agent whose nucleic acid sequence is known. The TemPCR method utilizes at least one set of target enrichment primers specific for the disease agent or secondary disease agent to be detected (present at a low concentration) and at least one pair of shared target amplification primers (present at high concentrations). At least one pair of said target enrichment primers comprises a binding sequence for the target amplification primers. Therefore, the use of the TemPCR method allows multiplex amplification reactions to be carried out without the need for empirical optimization of the multiplex amplification parameters.

Abstract: Disclosed is a method and composition of matter for PCR-based gene dosage analysis. The invention provides internal control DNA sequences that are the same length and same G-C content. The method does not require sized separation of the amplified products. Instead, the method utilizes hybridization and ELISA like colormetric screening. The invention further provides for tightly controlled internal standards for comparing gene dosage by placing one copy of various chromosome markers on one plasmid.