湖北地区汉族原发性先天性青光眼患儿CYP1B1基因突变分析

PCR-SSCP analysis of the CYP1B1 gene mutation in primary congenital glaucoma patients of Hubei Han nationality

目的研究湖北地区汉族原发性先天性青光眼(primary congenital glaucoma,PCG)患儿CYP1B1基因的突变情况。方法用苯酚-氯仿法从38例原发性先天性青光眼患儿的全血细胞中提取基因组DNA。然后通过聚合酶链反应-单链构象多态性(single-strand conformation polymorphism,PCR-SSCP)银染色法检测CYP1B1基因第2、3外显子的突变情况。结果5例PCG患儿检出CYP1B1基因第3外显子异常DNA片段条带,经测序分析证实为7990C→T,未发现第2外显子存在基因突变。结论CYP1B1基因具有明显的遗传异质性。应用PCR-SSCP技术可初步筛查原发性先天性青光眼患儿CYP1B1基因突变。

Objective To identify the CYP1B1 gene mutation in primary congenital glaucoma （PCG） patients of Hubeo Han nationality. Methods Genomic DNA was prepared from peripheral blood leucocytes of thirty-eight patients with PCG using phenol-chloroform extraction. CYP1B1 gene mutations （exons 2 and 3） were identified by silver staining of single-strand confomation polymorphism analysis of the polymerase chain reaction （PCR-SSCP）. Results Abnormal fragment bands of eron 3 were found in five cases of thirty-eight patient. Sequence analysis of those, abnormal DNA fragments demonstrated a C-to-T transition at base 7990. No eron 2 mutation was edtected. Conclusion Mutafions in the CYP1B1 gene have genetic heterogeneity. This indicates that PCR-SSCP may be. used to detect CYP1B1 gene mutation in preliminary tests.