摘要
目的探索新一代测序技术(NGS)在检测自然流产胚胎或绒毛组织染色体非整倍体和拷贝数变异(CNV)应用中的价值。方法选择20例自然流产患者的绒毛进行染色体核型分析,同时应用NGS技术进行染色体非整倍体和拷贝数变异的检测,并以染色体核型分析结果为"金标准"进行NGS方法的评估。后于2013年共收集1074例自然流产胚胎或绒毛组织,应用NGS技术完成染色体非整倍体和CNV的检测,并对检测结果进行分析。结果 20例自然流产样本的NGS结果与核型分析结果对比,检测灵敏度和特异性均为100%。临床检测的1074例流产组织样本中,42例样本DNA不符合质控标准,实际完成检测1032例。1032例组织样本中阳性445例(43.12%),其中非整倍体369例(82.92%),以16、X、22、21、15、18号染色体高发;CNV共76例(17.08%),阳性样本集中发生在8-12w。阴性587例(56.88%)。根据孕妇年龄将样本分为三组比较,差异有统计学意义(P<0.01)。结论 NGS技术用于检测流产组织的非整倍体和拷贝数变异具有较高的灵敏度和特异性,是适用于临床的有效检测方法。
Objective：To develop a novel method of testing aneuploidy and Copy number variation（CNV）in abortion fetus or chorionic villi using next generation sequencing（NGS）method,and to report the clinical application of this method in practice. Methods：We developed a NGS-based method to detect aneuploidy and CNV using low-coverage whole-genome sequencing and PSCC algorithm. To validate the method,a total of 20 cases of spontaneously abortion chorionic villi were tested by the NGS method,and compared to results of G-banding. And then,we collected 1074 samples which were abortion fetus or chorionic villi using NGS to detect the chromosome aneuploidies and CNV,and the test results were analyzed. Results：In the validation test,results from the NGS-based method in all 20 cases were consistent with the G-banding results,showing 100% of sensitivity and specificity. In clinical application of the NGS-based method,42 cases were failed to meet the sample quality criteria,ending with 1032 samples for valid testing. 369 cases with aneuploidy and 76 cases with CNV were identified,with the gestational week ranging from 8-12 weeks. Trisomy 16,X,22,21,15,18 were the most common aneuploidies. The testing positive samples showed a reduced occurrence rate as the gestational week grows,suggesting that spontaneous miscarriage due to aneuploidy and chromosome CNV is more likely to happen at early pregnancy. The samples is divided into three groups according to the pregnant women age,the difference was statistically significant（P0.01）. Conclusion：Our NGS-based method has a high sensitivity and specificity for detecting chromosome aneuploidy and CNV,so it has a clinical value for diagnosis of the genetic reasons of spontaneous miscarriage.
出处
《中国优生与遗传杂志》
2015年第2期12-15,共4页
Chinese Journal of Birth Health & Heredity
关键词
新一代测序技术
自然流产
绒毛膜绒毛
染色体畸变
Next generation sequence
Spontaneous miscarriage
Chorionic villi
Chromosome aberration
