孕妇血浆DNA测序在性连锁遗传病筛查中的应用

发布时间：2018-03-22 11:48

  本文选题：胎儿性别鉴定　切入点：孕妇血浆测序　出处：《华南理工大学》2014年硕士论文　论文类型：学位论文

【摘要】：目的——开发一种基于孕妇血浆测序的胎儿性别鉴定方法，评价其作为筛检试验的可能性，及其在性连锁遗传病咨询中的应用潜能。 方法——选取900例孕周为12-16周的、已知出生随访结果的孕妇血浆测序数据进行回顾性分析，其中训练集和验证集分别为100例和800例。统计比对到Y染色体上的测序片段的百分比，，基于ROC曲线方法选取性别鉴定分界值，并用于鉴定胎儿性别。招募8例杜氏肌营养不良症的携带者孕妇进行同步盲法测试，通过孕妇血浆测序，利用所建立的方法进行胎儿性别鉴定。对性别鉴定错误样本进行全面的回顾分析，包括常染色体数目异常检测及临床记录。其中，常染色体数目异常检测通过建立引入内部参考染色体的Z检验方法进行。 结果——在训练集中，本方法对胎儿性别鉴定的灵敏度和假阳性率分别为96%和0%。在验证集中，423例男胎中421例成功鉴别，377例女胎中374例成功鉴别，灵敏度及特异性达到99.53%和99.20%，其余各项评估指标均达到筛检试验要求。与羊水穿刺金标准对比，8例杜氏肌营养不良症携带者的胎儿性别鉴定均正确。引入内部参考染色体的Z检验方法对T13、T18和T21的检测灵敏度为100%，特异性为98.7%，100%和100%。使用该方法筛查所有5例性别鉴定错误样本，结果均为阴性。另外，该错误样本与孕周、孕妇年龄、唐氏风险和妊娠异常病史均没有直接关系。 结论——基于孕妇血浆测序，无创性的胎儿性别鉴定可达到高准确率。本方法在具有完善的伦理学考量的前提下，具有产前遗传咨询的应用前景。
[Abstract]:Objective to develop a method of fetal sex identification based on maternal plasma sequencing and to evaluate the possibility of screening test and its potential application in sex-linked genetic disease counseling. Methods the plasma sequencing data of 900 pregnant women aged 12-16 weeks with known birth follow-up results were retrospectively analyzed. The training set and validation set were 100 cases and 800 cases, respectively. The percentage of sequenced fragments on Y chromosome was statistically compared, and the gender identification boundary value was selected based on ROC curve method. It was used to identify fetal sex. Eight pregnant women with Duchenne muscular dystrophy were recruited for simultaneous blind test, and their plasma was sequenced. Using the established method for fetal sex identification. A comprehensive retrospective analysis of the wrong sex identification samples, including the detection of autosomal abnormalities and clinical records. The detection of autosomal abnormalities was carried out by establishing a Z test with internal reference chromosomes. Results in the training concentration, the sensitivity and false positive rate of this method for fetal sex identification were 96% and 0 respectively. Among 423 cases of male fetus, 421 cases were successfully identified from 377 cases of female fetus and 374 cases of female fetus were successfully identified. The sensitivity and specificity reached 99.53% and 99.20%, and the other evaluation indexes met the requirements of screening test. Compared with the gold standard of amniotic fluid puncture, the fetal sex identification of 8 cases with Duchenne muscular dystrophy was correct. The sensitivity and specificity of the Z method for the detection of T13, T18 and T21 were 100, 98.7% and 100%, respectively. The results were all negative. In addition, there was no direct relationship between the false sample and gestational age, age of pregnant women, Down's risk and history of abnormal pregnancy. Conclusion-based on maternal plasma sequencing, noninvasive fetal sex identification can achieve high accuracy. This method has the application prospect of prenatal genetic counseling on the premise of perfect ethical considerations.
【学位授予单位】：华南理工大学
【学位级别】：硕士
【学位授予年份】：2014
【分类号】：R714.5


本文编号：1648514