采用多重置换扩增进行脊肌萎缩症的植入前遗传学诊断研究  

Methodology study of preimplantation genetic diagnosis for spinal muscular atrophy by multiple displacement amplification

作　　者：沈晓婷[1] 张成[2] 徐艳文[1] 钟依平[1] 曾艳红[1] 王静[1] 丁晨晖[1] 周灿权[1] 

机构地区：[1]中山大学附属第一医院生殖医学中心,广州510080 [2]中山大学附属第一医院神经内科,广州510080

出　　处：《中国优生与遗传杂志》2015年第9期93-96,共4页Chinese Journal of Birth Health & Heredity

基　　金：广东省自然科学基金博士启动项目课题号:S2013040014613

摘　　要：目的建立一种可靠、准确的植入前遗传学诊断方法,可广泛用于各种类型的脊肌萎缩症。方法采用多重置换扩增(MDA)对单细胞进行全基因组扩增,等位基因特异性扩增SMN1外显子7进行脊肌萎缩症(SMA)致病基因检测及采用与SMN基因紧密连锁的12个短串联重复序列(STR)进行SMA的单体型分析。结果共进行了80单个淋巴细胞及63个单卵裂球的MDA。MDA扩增效率在单淋巴细胞中为97.5%(78/80),在单卵裂球为96.8%(61/63)。在单淋巴细胞及单卵裂球MDA产物中,总的PCR扩增效率和等位基因脱扣率(ADO)为分别为96.2%(587/610)和8.7%(52/597),95.5%(722/756)和9.9%(54/547)。在单淋巴细胞MDA产物中,SMN1外显子7的PCR结果均与外周血结果相符,而单个卵裂球MDA产物中,有一个PCR扩增失败,扩增效率为98.4%(60/61)。结论该方法可用于常见的缺失型的SMA的PGD,而且可以应用于突变型的为SMA的PGD,为进一步的临床应用奠定了基础。Objective：To establish a reliable and accurate prot℃ol in preimplantation genetic diagnosis（PGD）for spinal muscular atrophy（SMA）.Methods：The prot℃ol was comprised of single cell multiple displacement amplification（MDA）,allele-specific amplification of SMN1 exon7,singleplex fluorescent polymerase chain reaction（PCR）of twelve short tandem repeat（STR）markers flanking the SMN region. Results：MDA was successful in 97.5% of total 80 single lymph℃ytes and 96.8% of total 63 single blastomeres. PCR results of SMN1 exon7 with MDA products of single lymph℃ytes were in agreement with the results of peripheral blood test and the PCR efficiency of SMN1 exon7 for single blastomere was 95.2%（60/63）. The PCR efficiency of was 96.2%（587/610）for single lymph℃ytes compare to 95.5%（722/756）for single blastomeres. The ADO rate was 8.7%（52/597）for single lymph℃ytes and 9.9%（54/547）for single blastomeres,much lower compared with other reports on MDA using large amount of single cells. Conclusion：This method used mutation combine with haplotype analysis will improve PGD for SMA and can be used for all kinds of SMA.

关 键 词：脊肌萎缩症 植入前遗传学诊断 多重置换扩增 单体型分析 等位基因脱扣 

分 类 号：R440[医药卫生—诊断学] R714.8[医药卫生—临床医学]