应用SYBR Green I实时荧光定量聚合酶链反应检测常染色体隐性遗传早发性帕金森综合征的parkin基因外显子重排突变

Analysis of exon rearrangements in the parkin gene in patients with autosomal recessive early-onset parkinsonism using SYBR Green I Real-time PCR

目的建立应用SYBR Green I实时荧光定量聚合酶链反应（Real—time PCR，RT—PCR）检测parkin基因外显子重排突变的技术平台，应用该技术对常染色体隐性遗传早发型帕金森综合征（autosomal recessive early—onset parkinsonism，AREP）家系进行parkin基因外显子重排突变分析。方法应用SYBR Green I RT—PCR技术对32个中国AREP家系进行parkin基因外显子重排突变分析。结果14个家系先证者存在parkin基因外显子重排突变，其中3个为纯合缺失突变、3个为复杂杂合缺失突变和8个杂合缺失突变，未发现外显子重复突变，突变主要累及第2～4号外显子。结论建立了应用SYBR Green IRT-PCR技术检测parkin基因外显子重排突变的基因检测平台；中国AREP家系的parkin基因外显子重排突变频率为43．8％，与国外报道相似。

Objective To develop a method of detection exon rearrangements in the parkin gene （PARK2） using SYBR Green I real-time PCR and to analyze PARK2 exon rearrangement mutations in families with autosomal recessive early-onset parkinsonism （AREP） using this method. Methods Exon rearrangement in PARK2 was screened by SYBR Green I real-time PCR in 32 families with AREP. Results Exon rearrangement mutations were found in 14 families, including 3 compound heterozygous deletions; 3 homozygous deletions; and 8 heterozygous deletions. No duplication mutation was found. Hotspot for exon rearrangements clustered in exons 2 through 4. Conclusions We have developed a gene test method using SYBR Green I Real-time PCR to detect exon rearrangements in the gene PARK2. The frequency of PARK2 mutation is 43.8% in Chinese families with AREP. This frequency is similar to reported findings in other countries.