Analysis Of The CYP1B1 Gene Mutation In Primary Congenital Glaucoma Patients Of Hubei Han Nationality

[Objective] To identify the CYP1B1 gene mutation in Primary Congenital Glaucoma patients of Hubei Han nationality and help to establish the gene diagnosis of PCG.[Methods] Blood samples were collected from 47 patients with a diagnosis of PCG at Union Hospital and Children's Hospital. All of the patients and 100 unaffected members were studied. 1) Genomic DNA was extracted from the peripheral leukocytes of all subjects by the methods of phenol-chloroform extraction. 2) The primers were designed according to the references and two exons of the CYP1B1 gene were amplified using PCR method. 3) Gene mutations (exons 2 and 3) were identified by silver staining of single-strand conformation polymorphism technique and denaturing high performance liquid chromatograph (DHPLC). 4) Sequence analysis of the CYP1B1 gene.[Results] Two novel mutations were detected in 10 of the 47 unrelated patients. Direct sequencing demonstrated a homozygous C-to-T transition at codon 385 which produced L385F mutation and another A-to-G transition at codon 407 which produced T407A mutation of CYP1B1 gene[Conclusion] Mutation in exon 3 of CYP1B1 existed in PCG patients of Hubei Han nationality, and the novel mutations were probably pathological by nature. It is important that further study be conducted to seek for specific mutations of CYP1B1 gene or other possible relevant genes in PCG patients.