大规模并行测序用于胎儿染色体非整倍体的产前检测及羊水细胞染色体核型分析

发布时间：2018-06-27 16:42

本文选题：产前检测 + 羊膜腔穿刺术　；参考：《安徽医科大学》2014年硕士论文

【摘要】：目的应用大规模并行基因组测序技术和羊水细胞染色体核型分析,进行胎儿染色体异常的产前诊断;并探讨大规模并行基因组测序在产前检测中运用的可行性。方法选择2012年3月到2013年4月就诊,孕龄在16~23周之间高龄妊娠、唐氏综合征(Down syndrome)筛查高风险、不良生育史和B超显示胎儿异常等及无创DNA检测提示阳性孕妇共176例,抽取孕妇外周血,提取血浆DNA,制备文库,应用高通量基因测序仪检测,测得的基因序列与人类参考基因组比对并作统计分析;同时抽取羊水经细胞培养后行染色体核型分析。结果以羊水染色体核型分析结果作为金标准,对176例孕妇行大规模并行基因组测序。测序检出12例21三体型胎儿,164例非21三体型胎儿。其中12例21三体型胎儿中,经核型分析得到验证11例,1例测序结果为假阳性;测序方法对于21三体型胎儿的检出率为100%,特异性为99.39%,误诊率为0.61%,诊断符合率为99.43%。测序发现1例18三体型胎儿,175例非18三体型胎儿,与羊水染色体核型结果比对,测序方法对于18三体型胎儿的检出率为100%,特异性为100%,诊断符合率为100%。176例孕妇羊水染色体核型分析,检出异常核型23例,其中染色体数目异常11例,染色体结构异常12例。结论大规模并行基因组测序技术检测胎儿21、18号染色体非整倍体异常,其敏感性和特异性与染色体核型分析具有较高的一致性,且具有安全性高、准确性好、易于高通量检测等优势,值得临床推广。大规模并行基因组测序技术与羊水染色体核型分析的联合应用是保障母婴健康,减少出生缺陷的有效措施。
[Abstract]:Objective to study the feasibility of large-scale parallel genome sequencing and amniotic fluid cell chromosome karyotype analysis in prenatal diagnosis of fetal chromosomal abnormalities. Methods from March 2012 to April 2013, 176 pregnant women with gestational age between 16 and 23 weeks were selected for screening for Down syndrome, abnormal birth history, fetal abnormality and noninvasive DNA test. Maternal peripheral blood was extracted, plasma DNA was extracted and library was prepared. The sequence was compared with the human reference genome and analyzed statistically by high-throughput gene sequencer, and the chromosome karyotype was analyzed after the amniotic fluid was cultured. Results based on the results of amniotic fluid chromosome karyotype analysis, 176 pregnant women were sequenced. 12 cases of 21 trisomy fetus were sequenced and 164 cases of non 21 trisomy fetus were detected. Among 12 cases of 21 trisomy fetus, 11 cases were confirmed to be false positive by karyotype analysis, and the detection rate, specificity, misdiagnosis rate and diagnostic coincidence rate for 21 trisomy fetus were 100, 99.39, 0.61and 99.43g, respectively. The results of sequencing showed that one fetus with 18 trisomy and 175 non-18 triploid fetuses were compared with the results of amniotic fluid chromosome karyotype. The detection rate and specificity of 18 trisomy fetus were 100 and 100 respectively. The diagnostic coincidence rate was 100. 176 pregnant women with amniotic fluid chromosome karyotype analysis. 23 cases of abnormal karyotype were detected, including 11 cases of abnormal chromosome number and 12 cases of abnormal chromosome structure. Conclusion Large-scale parallel genomic sequencing technique for the detection of chromosomal aneuploidy abnormalities of fetal chromosomes 21 and 18 is highly consistent with chromosome karyotype analysis, and has high safety and accuracy. Easy to high throughput detection and other advantages, worthy of clinical promotion. The combined application of large-scale parallel genomic sequencing and amniotic fluid chromosome karyotype analysis is an effective measure to protect maternal and infant health and reduce birth defects.
【学位授予单位】：安徽医科大学
【学位级别】：硕士
【学位授予年份】：2014
【分类号】：R714.5

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