实时荧光定量PCR技术在年龄相关性白内障患者GSTM1、GSTT1基因拷贝数变异检测中的应用

发布时间：2018-10-12 17:40

【摘要】：目的应用实时荧光定量PCR技术检测年龄相关性白内障(age-related cataract,ARC)患者谷胱甘肽-S-转移酶(glutathione S transferase,GST)M1、T1基因拷贝数变异。方法采用实时荧光定量PCR技术检测279例(558眼)ARC组患者和145例(290眼)对照组的GSTM1、GSTT1基因重复或缺失等拷贝数变异情况,验证此方法在拷贝数检测中的准确性。结果相同条件下GSTM1、GSTT1基因和RNase P基因3次PCR扩增曲线基本重合、扩增效率基本一致、Ct值基本相同。GSTT1基因完全缺失(0个拷贝)的ARC个体(尤其是后囊下性ARC)和对照组相比,差异有统计学意义(P0.05);至少有1个拷贝缺失的GSTT1基因型的个体发生ARC和皮质性ARC的风险升高(OR值分别为2.16、4.81,均为P0.01),而2个拷贝缺失的GSTT1基因型的个体发生这种风险降低(OR=0.19,P0.05)。GSTM1基因拷贝数变异的ARC个体与对照组相比,差异无统计学意义(P0.05)。结论 GSTT1基因拷贝数的缺失可能是ARC特别是皮质性和后囊下性白内障发生的危险因素。实时荧光定量PCR检测准确、效率高,适用于GSTM1、GSTT1基因拷贝数变异的检测,适用于包括ARC在内的眼病流行病学调查研究。
[Abstract]:Objective to detect the copy number variation of glutathione-S-transferase (glutathione S transferase,GST) M _ 1 T _ 1 gene in patients with age-related cataract (age-related cataract,ARC) by real-time fluorescence quantitative PCR. Methods Real-time quantitative PCR was used to detect the GSTM1,GSTT1 gene repeat or deletion isocopy number variation in 279 patients (558 eyes) of ARC group and 145 cases (290 eyes) of control group. The accuracy of this method in the detection of copy number was verified. Results under the same conditions, the three PCR amplification curves of GSTM1,GSTT1 gene and RNase P gene basically coincided with each other, and the amplification efficiency was basically the same, and the Ct value was basically the same. The ARC individuals with complete GSTT1 gene deletion (0 copies) were compared with the control group (especially the posterior subcapsular ARC). The difference was statistically significant (P0.05); the risk of ARC and cortical ARC was increased in individuals with at least one copy deletion of GSTT1 genotype (OR = 2.16, 4.81, respectively, P 0.01), while in individuals with two copy deletion GSTT1 genotypes, the risk decreased. ARC individuals with low copy number variation of GSTM1 gene were compared with control group. The difference was not statistically significant (P0.05). Conclusion the loss of copy number of GSTT1 gene may be a risk factor for the occurrence of ARC, especially cortical and posterior subcapsular cataract. Real-time fluorescence quantitative PCR is accurate and efficient. It is suitable for the detection of copy number variation of GSTM1,GSTT1 gene and the epidemiological investigation of ophthalmopathy including ARC.
【作者单位】：南通大学附属医院眼科;
【基金】：国家自然科学基金资助(编号:81070718、81300744)~~
【分类号】：R776.1

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