基因芯片在卵巢组织HLA-DRB1分型中的应用

目的 用基因芯片对胎儿卵巢组织和成人卵巢组织进行HLA-DRB1分型,对于进一步了解胎儿卵巢基因型及其档案的建立进行了初步探索.方法 将6份胎儿卵巢组织分别与育龄妇女及绝经后妇女卵巢组织的基因组DNA,通过组间特异引物不对称扩增,扩增中同时用荧光素Cy3标记.扩增标记后的产物与结合在HLA-DRB1基因分型芯片上的探针进行杂交,通过荧江扫描仪对杂交产生的荧光信号值进行分析,确定样品的HLA-DRB1基因型.结果 18例卵巢组织样本中,纯合子只有1例,其余均为杂合子.其中两例胎儿卵巢组织的HLA-DRB1型别分别与一例育龄妇女卵巢组织及一例绝经后妇女卵巢组织相吻合.芯片重复率为100%.结论 基因芯片是一种理想的分型方法,具有特异性高、重复性好、操作简便、所需时间少、结果判读容易、一次可作多份样本的优点.通过这种技术对胎儿卵巢HLA-DRB1进行快速分型,可为卵巢移植手术的供受体选择节约了时间和成本,并可进一步建立胎儿卵巢组织的基因型档案,为卵巢移植工作开展奠定基础.

Genotyping of HLA-DRB1 by Oligonucleotide Array in Ovaries

Objective:To typing Human Leukocyte Antigen-DRB1(HLA-DRB1) alleles by oligonucleotide arrays in fetal ovaries and abult ovaries,and to develop the records of genotypes in fetal ovaries.Methods:Unsymmetrical PCR was used to amplify HLA-DRB1 gene,and the PCR products were labeled with Cy5.The labeled PCR products were used as templates for hybridization,then the signals were scanned by scanner and then analyzed and differentiate genotype by a set of computer software.Result:All the samples had been genotyped by HLA array successfully.And genetypes of two sample in fetal ovaries were respectively the same as two samples of adult ovaries.Conclusion: HLA-DRB1 genotyping by oligonucleotide array is a good method with advantage of high speed,low cost and high flux,and the genotyping chip is sensitive and specific and can test several samples at a time.Through the technique,it is more economical and quick to choose the appropriate donor and accepter in ovarian transplantation.,and to develop the records of genotypes in fetal ovaries.