Abstract: Nucleic acids can be amplified and detected using a very rapid polymerase chain reaction procedure. This procedure includes a series of steps which have critically defined temperature and time parameters. Each polymerase chain reaction cycle requires generally less than about two minutes, and in most cases less than 90 seconds. At least 5 units/100 &mgr;l of solution of thermostable DNA polymerase are used, and other preferred levels of primer concentrations facilitate the quick cycling in the amplification. In preferred embodiments, only two temperatures are used in the amplification.

Abstract: Nucleic acids can be amplified and detected using a very rapid polymerase chain reaction procedure in which two different nucleic acid sequences are present. This method allows one to preferentially modulate (for example, suppress) the degree of amplification of one or more nucleic acid sequences relative to other nucleic acid sequences. This modulation is achieved by exploiting differences in the relative primer melt temperatures, or by using certain ratios of primers. Each PCR cycle is very fast, that is less than about 90 seconds. This method is particularly useful for amplification and detection of DNA associated with infectious agents that may be present in a specimen in very small quantities compared to other nontargeted nucleic acids.