Dai Can, Li Wen, Gao Bodi, Li Lu-yun, Lu Guang-xiu
The Institute of Reproduction and Stem Cell Engineering, Xiangya Medical College, Central South University, People's Republic of China.
Zhonghua Yi Xue Yi Chuan Xue Za Zhi. 2008 Aug;25(4):373-7.
To identify the mutations of the tyrosinase gene (TYR) and P gene in patients with oculocutaneous albinism (OCA).
Polymerase chain reaction (PCR) and denaturing high performance liquid chromatography (DHPLC) were applied to detect the mutations in all exons of TYR gene and P gene. Then DNA sequencing and restriction endonuclease analysis were used to confirm the mutations detected by DHPLC. Novel mutations were screened in 100 unrelated persons with normal phenotypes to exclude the possibility of polymorphism.
Two mutations were detected in the P gene of the three patients and none in TYR gene. Heterozygous mutation of T450M in exon 13 of the P gene was detected in patient 1. Patient 2 had a heterozygous mutation of T450M in exon 13 and a heterozygous mutation of G775R in exon 23 of the P gene. Patient 3 had a heterozygous mutation of G775R as well. Restriction endonuclease analysis of the P gene exon 13 showed that the Oli I site had partly disappeared resulting from the heterozygous mutation T450M in patient 1 and patient 2, but not in 100 unrelated individuals. The heterozygous mutation T450M is a novel mutation.
Gene diagnosis of OCA can be carried out effectively by combining PCR, DHPLC, DNA sequencing and restriction endonuclease analysis.
鉴定眼皮肤白化病(OCA)患者酪氨酸酶基因(TYR)和P基因的突变情况。
应用聚合酶链反应(PCR)和变性高效液相色谱(DHPLC)检测TYR基因和P基因所有外显子的突变。然后采用DNA测序和限制性内切酶分析来确认DHPLC检测到的突变。在100名表型正常的无关个体中筛选新的突变,以排除多态性的可能性。
3例患者的P基因检测到2个突变,TYR基因未检测到突变。患者1在P基因第13外显子检测到T450M杂合突变。患者2在P基因第13外显子有T450M杂合突变,在第23外显子有G775R杂合突变。患者3也有G775R杂合突变。对P基因第13外显子进行限制性内切酶分析显示,患者1和患者2由于T450M杂合突变导致Oli I位点部分消失,而100名无关个体未出现这种情况。杂合突变T450M是一种新的突变。
联合应用PCR、DHPLC、DNA测序和限制性内切酶分析可有效进行OCA的基因诊断。