[Analysis of glucocorticoid receptor gene polymorphisms by denaturing high-performance liquid chromatography]

To detect polymorphisms of glucocorticoid receptor gene (NR3C1) in Chinese population with the technique of denaturing high-performance liquid chromatography (DHPLC).

Genomic DNA was isolated from peripheral blood of 2 healthy volunteer doctors and umbilical cord blood of 62 normal neonates. All the NR3C1-coding exons along with their flanking intron portions were amplified by polymerase chain reaction (PCR). For polymorphism screening, PCR products were analyzed by DHPLC. DNA fragments with aberrant elution profiles revealed by DHPLC were re-amplified and sequenced directly.

Eight polymorphisms were identified by DHPLC and sequencing analysis. Among them, 5 (1374A > G, IVSG-68_IVSG-63delAAAAAA, 2193T > G, IVSH-9C > G, 2382C > T) were novel and 3 (1896C > T, 2166C > T, 2430T > C) were reported previously. Two groups of polymorphisms ([1374A > G + IVSG-68_IVSG-63delAAAAAA + IVSH-9C > G + 2382C > T] and [1896C > T + 2166C > T + 2430T > C]) showed to have the complete linkage disequilibrium and were identified as 2 different haplotypes, which was first reported. The frequencies of the 2 novel haplotypes were 3.1% and 14.1%, respectively. Another polymorphism (2193T > G) was relatively rare.

DHPLC is a rapid and reliable method for polymorphism screening. Eight polymorphisms in the NR3C1 gene are detected in Chinese population with the technique of DHPLC, of which two haplotypes have been identified for the first time.