利用微卫星诊断早期自然流产绒毛组织染色体数目异常的研究  

Application of Microsatellite to the study of numerical chromosomal anomalies in early spontaneous miscarriages

作　　者：邹刚[1] 贺光[2] 陆建英[1] 骆敏[1] 郭茗[1] 杨慧珠[1] 段涛[1] 

机构地区：[1]同济大学附属第一妇婴保健院,上海200040 [2]上海交通大学BIO-X中心,上海200030

出　　处：《中国产前诊断杂志（电子版）》2008年第2期15-21,共7页Chinese Journal of Prenatal Diagnosis(Electronic Version)

基　　金：上海申康医院发展中心上海市级医院新兴前沿技术联合攻关项目(SHDC12006106)资助

摘　　要：目的运用微卫星(短串联重复序列,STR)结合的多重定量荧光PCR检测61例流产绒毛染色体数目异常,以评价它与传统经典的核型分析方法的关系。方法首先利用多重荧光定量PCR扩增48份无关个体DNA样本中的常见发生数目异常的染色体上19个STR位点和一个AMXY位点,扩增产物经毛细管电泳。计算出各位点的杂合度和正常杂合子峰面积比值范围。再对61例自然流产组织中这些位点进行扩增,判断染色体数目异常情况。染色体核型分析结果做为对照诊断标准。结果①核型分析成功率为65.6%;多重荧光定量PCR扩增成功率为98.3%。核型分析成功的40例样本,多重荧光定量PCR全部扩增成功,其中38例结果与核型分析结果相一致,以细胞遗传学作为诊断标准,诊断的符合率为95%。②运用核型分析方法,新鲜标本核型分析成功率为80.4%;不新鲜或者坏死标本的成功率为20%,P<0.001。运用STR-PCR方法,新鲜标本扩增成功率为100%;不新鲜或者坏死标本的成功率为93.3%,P=0.246。③AMXY和DXS8090、DXS8094联合诊断性别的成功率可达97.9%。结论结合STR的多重荧光定量PCR在诊断染色体数目异常中敏感性强,特异度高,可以做为细胞遗传学的补充。对于稽留流产和坏死变性的绒毛组织,STR-PCR也可以进行诊断。AMXY和DXS8090、DXS8094可用来进行性别诊断。Objective To incorporate the QF-PCR technique to the study of numerical chromosomal anomalies in miscarriage and evaluate its efficency with the classic method of karyotyping. Methods We amplified 2-3 microsatellites in each of the chromosome 13,14,15,16,18,21,22 and X in which aneuploids are commonly found.Capillary electrophoresis was used to analyze the PCR products.A trisomic sample can be diagnosed only if the marker was detected either as three peaks or as a pattern of two peaks with a ratio of>1.85.Results 65.6% of the 61 samples were successfully karyotyped,while the amplification success rate of the PCR was 98.3%.The corresponding rate of PCR to karyotype methods was 95%.Through karyotype method,the fresh sample group success rate was 80.4%,higher than 20% of the necrosis group,P<0.001,but in STR-PCR group,amplification rate of the two groups showed no difference(100% vs 93.3%,P=0.246).Genders of the 48 samples were correctly diagnosed by the trio-marker assay of AMXY,DXS8090 and DXS8094 except for the sample No.40.Conclusion QF-PCR represents a useful and reliable tool to diagnose numerical chromosomal anomalies.It also can provide a diagnosis even to missed abortion or necrosis tissue.AMXY combined with DXS8090 and DXS8094 can be used to perform sex determination.

关 键 词：自然流产 染色体核型分析 染色体数目异常 牙釉蛋白基因 定量荧光PCR 毛细管电泳 微卫星 

分 类 号：R714.21[医药卫生—妇产科学]