Diagnostic compositions, elements, methods and test kits for amplification and detection of human CMV DNA using primers having matched melting temperatures

This invention relates to diagnostic compositions, elements, methods and test kits for the amplification and detection of human cytomegaloviral (hCMV) DNA. In particular, it relates to improved methods of polymerase chain reaction (PCR) using test kits and buffered compositions containing "matched" primers for one or more target nucleic acids, one of which is hCMV DNA. An aqueous composition containing primers for opposing strands of human cytomegaloviral DNA and a second target DNA 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 Twithin the range of from about 65 to about 74° C., while the T's are within about 5° C. of each other. Such compositions are useful in diagnostic test kits and methods for amplification and detection of hCMV DNA and other target DNA's using multiple capture probes, in "multiplexing". All of the capture probes have T's which are greater than 50° C. and are within 15° C. of each other.