MiniSTR技术应用于孕妇血浆中胎儿游离DNA检测的研究  

Application of MiniSTR in the detection of fetal DNA in maternal plasma

作　　者：邓志辉[1] 李茜[1] 梁延连[1] 高素青[1] 邹红岩[1] 杨宝成[1] 

机构地区：[1]深圳市血液中心,广东深圳518035

出　　处：《中国输血杂志》2008年第7期498-501,共4页Chinese Journal of Blood Transfusion

基　　金：深圳市科技计划重点项目(编号200701014)

摘　　要：目的研究利用母血中胎儿游离DNA检测STR的方法。方法采集9名11—27孕周的孕妇抗凝血样并分离出血桨,提取血浆总DNA,利用ABI MiniFilerTM系统复合扩增D13S317、D7S820、D2S1338、D21S11、D16S539、D18S51、CSFIPO、FGA等8个常染色体miniSTR基因座和性别Amelogenin基因座,扩增产物经ABI 3100型基因测序仪作基因扫描检测,用GeneMapper ID 3.2软件分析结果。结果9例孕妇血浆样本中,均检出了父源性胎儿STR等位基因,D13S317等8个常染色体STR基因座中,平均检出胎儿特异等位基因3.1个/例。在性别Amelogenin基因座,Y等位基因检出的案例有5例,并且按ABI MiniFilerTM试剂盒的扩增条件,Amelogenin Y等位基因的荧光信号峰高均>50 RFU,平均占Amelogenin X等位基因峰高的8.45%;未检出Amelogenin Y等位基因的有4例,性别Amelogenin基因座的结果与出生后证实的胎儿性别相一致。结论MiniSTR技术适合于孕妇血浆中胎儿游离DNA的检测,在产前分子基因诊断中具有广泛的应用前景。Objective To develop an efficient method for detecting the short tandem repeat （ STR ） of fetal DNA in maternal plasma by miniSTR technique. Methods A total of 9 blood samples form pregnant women from 11 to 27 weeks of gestation were collected. Each isolated total plasma DNA was amplified in single multiplex using the ABI MiniFilerTM kit, which could simultaneously genotype the 9 miniSTR loci, including D13S317, D7S820, D2S1338, D21S11, D16S539, D18S51, CSFIPO, FGA and Amelogenin, and the PC R products were detected by using ABI PrismTM 3100 DNA Sequencer. The allelic designation of each STR locus was accomplished using the GeneMapper ID 3.2 software. Results Fatherorigin fetal STR allele was detected in all the 9 plasma DNA samples. An average of 3.1 fetal STR alleles of the 8 autosoreal STR loci was observed in each of the 9 plasma DNA samples. As for the Amelogenin locus, Amelogenin Y allele was detected in 5 plasma DNA samples from pregnancies with male fetus, and allelic peak height values were all over 50 RFU, according to ABI Mini FilerTM PCR conditions, and the ratio of Amelogenin Y allele peak height value to Amelogenin X allele peak height value was 8.45%. However, no Amelogenin Y allele was detected in other 4 plasma DNA samples from pregnant women with female fetus. Conclusion The miniSTR technique is suitable for STR genotyping using fetal DNA in maternal plasma, and it suggests a broad application in noninvasive molecular prenatal diagnosis.

关 键 词：短串联重复序列(STR) 短片段长度 MINISTR 胎儿游离DNA 产前诊断 

分 类 号：R445.112[医药卫生—影像医学与核医学] R714.52[医药卫生—诊断学]