Diagnostic compositions, elements, methods and test kits for amplification and detection of two or more target DNA's using primers having matched melting temperatures

摘要

An aqueous composition containing primers for opposing strands of two or more target nucleic acids can be used in polymerase chain reaction to provide simultaneously rapid and efficient amplification and detection of those nucleic acids. The primers for each target DNA differ in length by no more than 5 nucleotides and have a Tm within the range of from about 65 to about 74° C., while the Tm's are within about 5° C. of each other. Such compositions are useful in diagnostic test kits and methods for amplification and detection of multiple nucleic acids, or in "multiplexing", using multiple capture probes. All of the capture probes have Tm's which are greater than 50° C. and are within 15° C. of each other.