A Multiplex Polymerase Chain Reaction Method for the Simultaneous Detection of GSTM1, GSTT1, and GSTP1 Polymorphisms

Abstract

Glutathione S-transferases (GSTs) play an important role in the detoxification of a wide variety of toxicants. Among GSTs, GSTM1 and GSTT1 polymorphic deletions and the GSTP1 Ile105Val polymorphism were often studied in combination in many diseases because of their additive effects, but they were usually genotyped by two separate methods. The purpose of the present study was to develop a simple and reliable method to simultaneously detect these three polymorphisms. The three polymorphisms of 259 volunteers were genotyped using a multiplex polymerase chain reaction (PCR) method based on a tetra-primer amplification refractory mutation system-PCR (T-ARMS-PCR), and the results were validated by multiplex PCR for GSTM1 and GSTT1 polymorphisms and DNA sequencing of the GSTP1 Ile105Val polymorphism, respectively. The multiplex PCR method of GSTM1, GSTT1, and GSTP1 polymorphisms based on T-ARMS-PCR can simultaneous detect the three polymorphisms in a single PCR. The results of this method are in perfect accord with the results of the multiplex PCR method of determining GSTM1, GSTT1 polymorphisms, and DNA sequencing of GSTP1 Ile105Val polymorphism. The novel multiplex PCR method of GSTM1, GSTT1, and GSTP1 polymorphisms is simple, fast, low-cost, and reliable for the simultaneous detection of GSTM1, GSTT1, and GSTP1 Ile105Val polymorphisms.