PARK16与苏州地区汉族人帕金森病相关性研究

Association of PARK 16 polymorphisms with Parkinson＇s disease in Han population of Suzhou area

目的 研究PARK16区域rs947211多态性与帕金森病（PD）的关系.方法 采用病例-对照研究,病例组为2008至2010年之间就诊于苏州大学附属第二医院帕金森病专科门诊的160例PD患者,对照组为来自我院体检中心的200名健康正常人,采用聚合酶链反应（PCR）、DNA测序、高保真DNA聚合酶介导等位基因特异性PCR结合实时荧光PCR的SNP检测方法,检测PARK16区域rs947211多态性,探讨其与苏州地区PD发病的关系.结果 成功建立了高保真酶介导的rs947211多态性检测方法,并得到DNA测序结果验证,采用荧光检测提高了检测效率和通量.研究发现了PARK16区域rs947211的所有基因型,PD组G等位基因频率高于对照组（60.6%比51.0%）,且差异有统计学意义（χ2=6.662,P=0.010,OR=0.676,95%CI0.502～0.911）,两组中G/G基因型与A/G＋A/A基因型相比差异有统计学意义（χ2=8.401,P=0.004,OR=1.951,95%CI 1.238～3.076）.H＆Y分级在不同基因型之间差异有统计学意义（P＜0.05）,G/G基因型分级明显高于A/A基因型分级（P=0.025,95%CI0.059～0.851）和A/G基因型（P=0.020,95%CI 0.058～0.654）.结论 结合高保真酶介导的等位基因SNP识别方法建立了高效特异的实时荧光PCR检测rs947211多态性的方法.多态性检测结果提示苏州地区汉族人群中PARK16区域与PD相关,证实了对日本PD患者的全基因组扫描的研究结果.另外,对临床特点的分析发现,G/G基因型可能是PD的危险因素,在3个基因型中,G/G基因型携带者的疾病严重度或许最高.

Objective To investigate whether there is an association of PARK 16 locus with Parkinson＇s disease （PD） in Han population of Suzhou area. Methods Polymerase chain reaction （PCR）, sequencing analysis, allele-specific PCR mediated by pfu DNA polymerases and real-time PGR were employed to determine the genotype of each subject. The PARK16 representative SNP rs947211 was analyzed in 160 PD patients and 200 age-matched healthy controls. Results The results of pfu-mediated allelespecific PGR were 100% concordant with the direct-sequencing analysis. All genotypes were found in this study. A significant distinction occurred in the allelic frequency of rs947211. There was a higher expression of G allele in PD patients versus the controls （60. 63% vs 51.00% χ2 =6. 662, P =0. 010, OR =0. 676,95% CI0. 502 - 0. 911 ）. There were remarkable differences between G/G genotype and A/G ＋ A/A genotype of two groups（χ2 =8. 401, P = 0. 004, OR = 1. 951, 95% CI 1. 238 - 3. 076）. In addition, a significant distinction also existed in different genotypes through H＆Y stage （P 〈 0.05）. For example, the stage of G/G was higher than A/A genotype （ P = 0. 025,95% CI 0. 059 - 0. 851 ） or A/G genotype （ P =0. 020,95% CI 0. 058 -0. 654 ）. Conclusion We have established a simple and reliable rs947211genotyping method mediated by pfu DNA polymerases. The results confirmed the existence of PARK16 in Han population of Suzhou area, rs947211 G/G genotype maybe a risk factor for PD and G/G genotype carriers perhaps obtain higher illness severity.