摘要
目的研究2个无关的A型尼曼-匹克病家系酸性神经鞘磷脂酶(ASM)基因SMPD1突变及遗传特征,分析基因型与表型关系。方法收集2位先证者及其家庭成员的临床资料,采用基因组小剂量抽提试剂盒从2个家系共6名成员及100例健康对照者外周血中提取基因组DNA,根据人类基因组数据库中获得的基因序列(NM000543)设计4对引物,采用聚合酶链反应(PCR)进行扩增,并应用PCR产物回收试剂盒进行产物回收,采用DNA直接测序法进行基因突变检测,测序结果应用DNAman软件进行序列对比分析,确定基因突变位点,对测序异常的片段重新进行PCR扩增与测序,以验证结果的可靠性。结果 PCR扩增所得片段与预计扩增片段大小一致,无非特异扩增带,可以进行纯化与测序。将测序结果与正常序列进行对比分析后,在先证者1和2的SMPD1基因的第1号外显子均发现一个纯合突变T107C,遗传密码子由GTG变为GCG,从而导致36位缬氨酸被丙氨酸替代(p.V36A),最终致使ASM(E.C.3.1.4.12)活性缺陷,产生临床症状。先证者之父母为携带者,100例健康对照中未发现上述突变。结论证实SMPD1基因是本研究中2个无关的A型尼曼-匹克病家系的致病基因。2个家系患儿均为SMPD1基因纯合突变致病,其父母为表型正常的基因突变携带者。
Objective To study the acid sphingomyelinase（ASM） gene（SMPD1）mutation in 2 unrelated Chinese families with type A Niemann-Pick Disease（NPD） and analyze the phenotype and genotype relationship.Methods Two NPD families after patients or parental consent were collected.Genomic DNA samples were extracted from peripheral bloods of the probands,their family members and 100 unrelated healthy individuals.Analysis of 6 exons and part exon-intron boundaries of the SMPD1 gene in patients,their parents and healthy individuals were carried out by polymerase chain reaction and direct DNA sequencing.Results The PCR amplification clips which were consistent with the expected one could be purified and sequenced.One homozygote mutation of SMPD1,which named as T107C（V36A） was identified both in 2 probands.Parents of the 2 probands had the hetemzygous mutation（c.107T→C,p.V36A） in exon 1 while 100 healthy individuals had no mutation.Conclusions The SMPD1 gene is related with the clinical phenotype in 2 Chinese NPD patients who are homozygote mutations.Their parents are carrier with normal phenotype.
出处
《实用儿科临床杂志》
CAS
CSCD
北大核心
2011年第12期952-954,958,共4页
Journal of Applied Clinical Pediatrics
