紧密连锁的多态性位点在β地中海贫血植入前遗传学诊断中的应用(英文)  

Closely linked polymorphic marker: successful application in preimplantation genetic diagnosis for beta-thalassemia

作　　者：邓捷[1] 庄广伦[1] 彭文林[1] 周灿权[1] 李洁[1] 方丛[1] 李穗萍[1] 陈勇[2] 童大跃[2] 

机构地区：[1]中山大学附属第一医院生殖中心,广州510080 [2]中山大学附属第一医院法医学教研室,广州510080

出　　处：《中华医学遗传学杂志》2005年第4期391-395,共5页Chinese Journal of Medical Genetics

基　　金：卫生部临床学科重点资助项目(2001321)~~

摘　　要：目的探讨与β珠蛋白基因紧密连锁的多态性位点HumTH01在β地中海贫血(β地贫)植入前遗传学诊断(preimplantationgeneticdiagnosis,PGD)中的作用。方法对4例已出生重型β地贫患儿的、双方均为β地贫基因携带者的夫妇进行了6个周期的PGD治疗,应用多重巢式PCR同时检测β珠蛋白基因及HumTH01基因,选择健康的胚胎移植入子宫。结果6个周期共活检44个胚胎,获得44个卵裂球,其中41个卵裂球扩增成功,35个胚胎经PCR分析后获得明确诊断,移植了14个胚胎,获得1例临床妊娠。孕17周时经脐带血穿刺,证实为完全正常胚胎,现已出生一正常女婴。单个卵裂球平均扩增效率为89.7%,等位基因脱扣(alleledropout,ADO)率为14.4%。HumTH01基因可以帮助检测出ADO及污染的发生。结论本研究为国内首次报道应用多重巢式PCR同时检测β珠蛋白基因及HumTH01基因对β地贫进行植入前遗传学诊断并成功获得临床妊娠。在PGD中同时检测与β珠蛋白基因紧密连锁的多态性位点可以降低PGD中由于ADO及污染造成的误诊的风险。Objective To evaluate the applicability of the polymorphic marker closely linked with beta-globin gene for the preimplantation genetic diagnosis （PGD） in couples at risk of having child with beta-thalassemia. Methods Single cell multiplex nested PCR which coamplifies the beta-globin gene and the closely linked polymorphic marker, HumTHO1 gene, was applied in six clinical PGD cycles for four couples with beta-thalassemia. Results In six clinical PGD cycles, a total of 44 embryos were biopsied and 44 blastomeres were obtained. Forty-one blastomeres were amplified and thirty-five embryos were given definite diagnoses. Fourteen embryos were transferred back to the uterus of the patients and one pregnancy went on well and ended with one live healthy birth, which confirmed the results of PGD. The average amplification efficiency of single blastomere was 89.7% and the average allele drop-out（ADO） rate was 14.4%. The coamplification of HumTHO1 could help to detect the existence of ADO and contamination. Conchusion This is the first report on unaffected pregnancy resulting from PGD using multiplex nested PCR in China. The simultaneous amplification of polymorphic marker closely linked to beta-globin gene （HumTHO1） could help to resist the risk of misdiagnosis in PGD caused by ADO and contamination.

关 键 词：基因多态性 Β地中海贫血 植入前遗传学 诊断方法 基因脱扣 

分 类 号：R446.9[医药卫生—诊断学]