MULTIPLEX POLYMERASE CHAIN-REACTION FOR HUMAN HERPESVIRUS-6, HUMAN CYTOMEGALOVIRUS, AND HUMAN BETA-GLOBIN DNA

Human cytomegalovirus and human herpesvirus-6 are closely related viru ses which cause similar diseases, have similar cellular repositories o f latent infection, and may be detected largely in the same types of c linical specimens. DNA amplification appears likely to play an increas ing role in the diagnosis of recent and remote infection with these ag ents. A sensitive multiplex polymerase chain reaction was therefore de veloped for the two viruses and for human P-globin DNA. Optimization o f parameters such as the primers, primer concentrations, magnesium con centration, and buffer constituents was crucial in achieving a sensiti ve assay. Preliminary results indicated that the assay could simultane ously monitor DNA extraction from clinical specimens and allow detecti on of HCMV or HHV-6 in patients with diseases possibly caused by eithe r pathogen.