Multiplex PCR and multicolor probes melting for the simultaneous detection of five UGT1A1 variants

The multiplex PCR melting analysis method was developed for detecting the five UGT1A1 variants. Multiplexing was achieved using color probes and T_m. The probes for *28/*6, *27, *29, and *7 were discriminated by colors. Although the probes for *28 and *6 had the same colors, their variants were clearly discriminated by probe T_m. The allelic frequencies of each genotype were 0.12 for *28, 0.19 for *6, 0.02 for *27, 0.0 for *29, and 0.005 for *7. We developed a multiplex PCR melting analysis method, which will be useful in molecular diagnostics and pharmacogenetic analyses in clinical laboratories.