High-throughput genotyping method for glutathione S-transferase T1 and M1 gene deletions using TaqMan (R) probes

A high-throughput genotyping method has been developed to detect gene delet ion polymorphisms of glutathione-S-transferase theta and mu (GSTT1 and GSTM 1). This method utilizes the 5'-nuclease activity of Tag polymerase in conj unction with fluorogenic TaqMan(R) probes. In contrast to traditional allel ic discrimination genotyping to detect single nucleotide polymorphisms, the current assay has been designed to detect gene deletion by utilizing custo m-designed TaqMan probes in conjunction with an exogenous internal positive control probe. The TaqMan genotyping results were validated by a commonly used multiplex PCR technique. Screening of 71 unrelated individuals reveale d gene deletion (null) genotype of 15.5% and 40.8% for GSTT1 and GSTM1, res pectively. This TaqMan genotyping method is rapid, reproducible, and highly sensitive and could be applied toward fully automated large-scale genotypi ng.