胚胎植入前地中海贫血检测试剂评价

Evaluation of preimplantation genetic diagnosis reagent for thalassemia

目的使用胚胎植入前地中海贫血诊断国家参考品,评价基于半导体测序法的胚胎植入前地中海贫血检测试剂的性能。方法将家系中模拟胚胎细胞样本进行单细胞全基因组扩增。取家系中其它样本和胚胎细胞的基因组DNA作为模板,扩增选定区域,加上通用的测序接头完成文库构建;将文库进行定量;使用测序仪进行测序。结果在国家参考品1号家系中,胚胎CNGB030013的β珠蛋白基因(HBB)基因检测结果:单体型结果为M1/F2(即遗传CNGB030012女方风险染色体中国型δβ,CNGB030011男方正常染色体β^(N));其致病基因检测结果为中国型δβ/β^(N)。HBA基因的检测结果:单倍体结果为M2/F1(即遗传CNGB030012女方正常染色体αα,CNGB030011男方风险染色体α^(⁃3.7)),其致病基因检测结果为αα/α^(⁃3.7)。在国家参考品2号家系中,胚胎CNGB030017的HBA基因检测结果:单体型结果为M1/F2(即遗传CNGB030016女方风险染色体αα^(CS),CNGB030015男方正常染色体αα);其致病基因检测结果为αα^(CS)/αα。在国家参考品3号家系中,胚胎CNGB030021的HBB基因检测结果:单体型结果为M2/F1(即遗传CNGB030020女方正常染色体β^(N),CNGB030019男方风险染色体中国型δβ);其致病基因检测结果为β^(N)/中国型δβ。在国家参考品4号家系中,胚胎CNGB030010的HBB基因检测结果:单体型结果为M2/F1(即遗传CNGB030008男方风险染色体β^(IVS⁃II⁃654),CNGB030009女方正常染色体β^(N));其致病基因检测结果为β^(N)/β^(IVS⁃II⁃654)。结论研制的国家参考品能够满足胚胎植入前地中海贫血检测试剂质量评价的要求。

Objective To evaluate the performance of preimplantation genetic diagnosis reagent for thalassemia based on semiconductor sequencing using national reference for preimplantation thalassemia diagnosis.Methods DNA was extracted from simulated embryos cells in the pedigree by whole genome amplification of single cell.The genomic DNA from other samples in the pedigree and embryos cells were used as templates to amplify selected regions,and common sequencing connectors were added to complete library construction.Then the library was quantified.Finally,sequencer was used for sequencing.Results In the national reference family No.1,the HBB gene test result of embryo CNGB030013:the haplotype result is M1/F2(that is,the CNGB030012 female risk chromosome Chinese typeδβ,the CNGB030011 male normal chromosomeβ^(N));the causative gene test result is Chinese typeδβ/β^(N).HBA gene test result:the haploid result is M2/F1(ie,the CNGB030012 female normal chromosomeαα,the CNGB030011 male risk chromosomeα^(⁃3.7)),and the causative gene test result isαα/α^(⁃3.7).In the national reference family No.2,the HBA gene test result of embryo CNGB030017:the haplotype result is M1/F2(that is,the CNGB030016 female:risk chromosomeαα^(CS),the CNGB030015 male normal chromosomeαα);the causative gene test result isαα^(CS)/αα.In the national reference family No.3,the HBB gene test result of embryo CNGB030021:the haplotype result is M2/F1(that is,the CNGB030020 female normal chromosomeβ^(N),the CNGB030019 male risk chromosome Chinese typeδβ);the causative gene test result isβ^(N)/Chinese typeδβ.In the national reference family No.4,the HBB gene test result of embryo CNGB030010:the haplotype result is M2/F1(that is,the inheritance of CNGB030008 male risk chromosomeβ^(IVS⁃II⁃654,)CNGB030009 female normal chromosomeβ^(N));the causative gene test result isβ^(N)/β^(IVS⁃II⁃654).Conclusion The developed national reference material can meet the requirements of quality evaluation of preimplantation genetic diagnosis reagent for thalassemia.