Multiplex-PCR was first described in 1988 as a method to detect deletions in the dystrophin gene.
[3] In 2020, RT-PCR multiplex assays were designed that combined multiple gene targets from the Center for Diseases and Control in a single reaction to increase molecular testing accessibility and throughput for SARS-CoV-2 diagnostics.
[4] Multiplex-PCR consists of multiple primer sets within a single PCR mixture to produce amplicons of varying sizes that are specific to different DNA sequences.
Annealing temperatures for each of the primer sets must be optimized to work correctly within a single reaction, and amplicon sizes, i.e., their base pair length, should be different enough to form distinct bands when visualized by gel electrophoresis.
Commercial multiplexing kits for PCR are available and used by many forensic laboratories to amplify degraded DNA samples.