Whole exome sequencing identifies a novel FRAS1 mutation and aids in vitro fertilization with preimplantation genetic diagnosis in Fraser syndrome

ObjectiveTo demonstrate the picture of a woman who had three times of pregnancies but fetuses were complicated with Fraser syndrome, a rare genetic disorder with multiple congenital anomalies.Case reportHere are three complicated pregnancies with predominant features of severe oligohydramnios and other variable intrafamilial presentations. We made a definite diagnosis, Fraser syndrome, with the assistance of whole exome sequencing (WES) via umbilical blood of the second and third fetus. The provision of a preimplantation diagnosis helped contribute a healthy newborn in this family.ConclusionThis paper provides insights into obscure antenatal presentations of Fraser syndrome with intrafamilial variance. Clinical uncertainty at the fetal stage suggests the role of WES to reach a final diagnosis, and a preimplantation diagnosis is applicable to avoid recurrence of genetic disorders in subsequent pregnancies.     

Routine screening with fetal echocardiography for prenatal diagnosis of congenital heart disease

Congenital cardiac anomalies are the most common congenital anomalies, occurring in approximately eight of 1000 live births. Proper perinatal and neonatal management is dependent upon accurate prenatal diagnosis. Approximately 10% of fetuses with cardiac abnormalities have identified risk factors; hence, most of the anomalies occur in pregnancies without prenatal risk factors. The application of detailed fetal echocardiography for prenatal screening, at present reserved mainly for high-risk cases, requires further evaluation before being recommended for the general population.

This article presents our experience of evaluating the accuracy of fetal echocardiography as a screening method in detecting cardiac anomalies in the general population of Singapore. We reviewed data from 39 808 pregnant women who received antenatal care at the National University Hospital, Singapore, between January 1986 and December 1994, and who underwent routine fetal echocardiography at 21-22 weeks of gestation. We identified 294 cases of congenital heart defects by fetal echocardiography. We obtained a sensitivity of 85.4% for the detection of congenital heart disease, and a specificity of 99.9% to rule out such anomalies. Our positive and negative predictive rates were 87.7% and 99.9%, respectively.

We recommend routine screening by echocardiography of all pregnancies at 21-22 weeks of gestation, irrespective of risk stratifcation, for the prenatal detection of cardiac anomalies, in order to improve perinatal management.     

实时荧光定量聚合酶链反应技术在产前诊断唐氏综合征中的应用

目的探讨实时荧光定量PCR技术用于产前诊断唐氏综合征的可行性。方法采用实时荧光定量PCR法,分别检测85例唐氏综合征高风险的中期妊娠妇女的羊水和7例智残儿外周血标本中,21号染色体上特异区域基因片段(DSCR3)和管家基因片段(GAPDH),并计算两者的比值。同时采用细胞遗传学方法分析其染色体核型。结果80例羊水细胞DNA检测DSCR3／GAPDH比值在0．46—1．30之间,染色体核型全部正常;而另5例羊水细胞和7例智残儿外周血DSCR,／GAPDH比值明显升高,达1．64～1．98,染色体核型均为21三体。5例中有3例核型为47,XY, 21;1例核型为47,XX, 21;另1例为易位型[46,XY,-15, t(15;21)]。7例智残儿中4例为47,XY 21;3例为47,XX 21。结论实时荧光定量PCR技术可作为产前快速准确诊断唐氏综合征的有效方法。     

改良羊水原位培养法进行产前诊断的研究

目的 :探讨改良羊水原位培养法在产前诊断中的价值。方法 :抽取 138例孕16～ 30周孕妇的羊水 ,用全营养培养基 4ml+羊水细胞混悬液 5ml接种培养 ,适时改良法原位收获制片、分析。结果 :138例羊水培养均获成功 ,成功率 10 0 % ;平均培养时间为 7天 ;平均可分析核型数 38个 ,发现染色体异常 4例。结论 :用此法培养羊水成功率高 ,培养时间短 ,可分析核型多 ,培养适用范围宽 ,可满足临床产前诊断的要求     

A novel nonsense mutation of TGFBR1 in a fetus with untypical Loeys-Dietz syndrome 1

ObjectiveWe present a rare untypical Loeys-Dietz syndrome 1 case in prenatal setting and report a novel mutation in the TGFBR1 gene.Case reportA pregnant woman came for medical attention due to the fetal ultrasound anomaly. The fetus was found to have short long bones. Trio-based WES was applied to the family. A novel de novo nonsense mutation c.1237C > T was detected in the TGFBR1 gene. A diagnosis of Loeys-Dietz syndrome 1 (LDS1) was plausible, but the fetus did not demonstrate the characteristic phenotype of the syndrome.ConclusionIn prenatal setting, fetal phenotypes are difficult to be fully observed, putting stress on the utility of molecular techniques. LDS1 in fetuses could present untypical features such as skeletal dysplasia.     

The outcome of gastroschisis after a prenatal diagnosis or a diagnosis only at birth. Recommendations for prenatal surveillance

OBJECTIVES: To establish in infants with gastroschisis whether outcome is different when comparing a prenatal diagnosis with a diagnosis only at birth with the intention to develop a prenatal surveillance protocol. Intestinal atresia established after birth and preterm versus term delivery were studied as risk factors. STUDY DESIGN: All 24 fetuses and 9 infants diagnosed with gastroschisis and referred to our tertiary center between January 1991 and June 2003 were studied retrospectively. RESULTS: The infants of the prenatal subset delivered at our tertiary center and 18 survived. There were two pregnancy terminations, three intrauterine deaths at 19, 33 and 36 weeks respectively and one neonatal death. All nine infants in the postnatal subset survived. Eight were out born and one was delivered at our tertiary center. Prenatal bowel dilatation did not correlate with outcome. Between the prenatal and postnatal subset no significant difference in outcome of live-born infants was established. For four infants with intestinal atresia a significant difference was demonstrated for induction of preterm labour (P<0.05), duration of parenteral nutrition (P<0.01), number of additional surgical procedures (P<0.001) and length of hospital stay (P<0.01). The fifteen infants born prior to 37 weeks of gestation spent a significantly longer period in hospital compared to those delivered at term. When the cases with bowel atresia were excluded this difference was no longer present. Five of the 33 cases were diagnosed with associated anomalies which mainly involved the urinary tract. CONCLUSION: Neonatal outcome of live born infants following a prenatal diagnosis of gastroschisis is not different from a diagnosis at birth. The presence of intestinal atresia is the most important prognostic factor for morbidity. The supplemental value of prenatal diagnosis to the outcome of infants with gastroschisis may be in the prevention of unnecessary intrauterine death and detection of intestinal complications. A proposed surveillance protocol for fetuses with gastroschisis focused on intrauterine signs of pending distress such as a dilated stomach, intra abdominal bowel dilatation with peristalsis, notches in the umbilical artery Doppler signal, development of polyhydramnios and an abnormal CTG registration may improve outcome.     

荧光定量PCR技术用于行遗传性耳聋基因检测胎儿的21三体综合征筛查的可行性

目的 探讨荧光定量PCR(QF-PCR)技术用于产前行遗传性耳聋基因检测的胎儿筛查21三体综合征的可行性.方法 选择2009年3月至2010年5月在解放军总医院行产前遗传性耳聋基因检测的54例孕妇为研究对象.所有行遗传性耳聋基因检测孕妇的家庭中的先证者均经过解放军总医院耳鼻咽喉研究所聋病分子诊断中心明确了耳聋致病基因.54例孕妇平均年龄31岁,其中9例≥35岁;孕周为18～26周,其中＞23孕周的孕妇有5例.孕18～23周的49例孕妇取羊水15～20 ml进行胎儿耳聋基因检测,孕周＞23周的5例,取新生儿脐带血1～2 ml进行相关耳聋基因检测.年龄≥35岁的9例孕妇中,5例同时进行了羊水细胞培养和染色体核型分析.采用QF-PCR技术诊断21三体综合征,共选择21号染色体杂合度高的9种短串联重复序列(STR)分子标记,选定6对引物分两种组合进行多重扩增,21三体综合征诊断标准为21号染色体2个以上STR位点出现1∶1∶1等比例峰及2∶1比例峰,或1∶2比例峰,即可明确诊断.结果 (1)遗传性耳聋基因的检测及其随访结果:54例胎儿均成功进行了相应的耳聋基因(包括GJB2基因、SLC26A4基因等)检测.10例胎儿的耳聋基因与先证者的基因类型相同,选择终止妊娠;其余44例没有重复先证者的基因类型,临床预测不会罹患由先证者基因改变引起的耳聋,选择继续妊娠,现均已分娩,新生儿出生后由专人随访,检查其听力均正常.(2)QF-PCR技术诊断21三体综合征的结果:54例胎儿全部检测成功,两对组合引物同时扩增后,均明确排除21三体综合征的诊断.5例高龄孕妇同时进行了胎儿染色体核型分析也均未见异常.新生儿出生后随访其发育情况,从外观也排除了21三体综合征的诊断.QF-PCR技术检测结果在1～3 d即可获得,无漏诊及误诊.结论 对于产前行遗传性耳聋基因诊断及其他产前基因诊断的胎儿,同时行QF-PCR技术检测可明确胎儿是否为21三体综合征;QF-PCR技术检测21三体综合征具有快速、准确、价格低廉及高通量等优点,且不增加羊水及脐血标本的抽取量.

Abstract：

Objective To establish the genetic test technique of trisomy 21 concurrently conducts with prenatal diagnosis for hereditary hearing loss. Methods Fifty-four pregnant women who underwent prenatal diagnosis for hearing loss of their fetuses in Chinese People's Liberation Army General Hospital from March 2009 to May 2010 were enrolled in this study. All probands from the deaf families have confirmed the causative mutation for hearing loss in Genetic Testing Center in Chinese People's Liberation Army General Hospital. The mean age of 54 pregnant women is 31 years at pregnancy of 18 - 26 weeks, 5 cases ＞ pregnancy of 23 weeks, 9 cases ≥ 35 years. All subjects did not conduct the serologic tests for trisomy 21before. Fifteen to twenty ml amniotic fluid was drawn from 49 cases at pregnancy of 18 - 23 weeks and 5 cases ＞ pregnancy of 23 weeks. One to two ml umbilical blood was drawn from 5 cases ＞ pregnancy of 23 weeks. For 9 cases ≥ 35 years, amniotic fluid cell culture and karyotyping analysis were conducted concurrently. A multiple quantitative fluorescent ( QF) PCR and six microsatellite markers were applied to as trisomy 21. Results (1) Fifty-four fetuses were successfully conducted prenatal genetic diagnosis for hearing loss (included GJB2 and SLC26A4). Ten fetuses copied the exactly same genotypes as the probands. The other 44 cases fetuses did not copy the same genotypes as the probands and won't develop hearing loss. The hearing test showed normal hearing for the neonates. (2) All the 54 fetuses were excluded of trisomy 21 by QF-PCR and were verified after birth. Five fetuses with advanced maternal age were performed karyotyping analysis and showed normal. The diagnostic results of QF-PCR can be obtained in 1 - 3 days without misdiagnosed. Conclusions QF-PCR is an efficient, rapid and accurate technique for detection of trisomy 21 without increasing sample amount. It can be used for fetuses who were undertaken hearing loss gene test or other prenatal gene test.     

高龄孕妇对2种不同唐氏产前诊断方案的需求及经济学评价

目的:探讨高龄孕妇对2种不同唐氏产前诊断方案的需求,并对这2种方案进行经济学评价。方法:选择2015年3月1日至2016年3月31日孕20+6周前在深圳市妇幼保健院产科就诊的单胎高龄孕妇4104例,由孕妇自主选择先行无创胎儿DNA检测(NIPT),NIPT高风险再行羊膜腔穿刺(方案1)或直接羊膜腔穿刺(方案2)。比较孕妇对2种方案的需求及其影响因素,并对2种方案进行经济学评价。结果:选择方案1的比率明显高于方案2[51.34%(2107/4104)vs 7.65%(314/4104),χ~2=1883.431,P0.001]。Logistic回归分析显示,孕妇年龄、传统唐氏筛查结果和胎儿颈部透明层厚度(NT)是影响孕妇选择的因素。与方案2相比,方案1可以节省67.21%的总费用。结论:高龄孕妇对NIPT有巨大的需求,且高龄孕妇采用NIPT筛查阳性后再行羊膜腔穿刺术可节省医疗费用,建议在充分知情同意的基础上可将NIPT用于高龄孕妇的一线筛查,尤其是适用于年龄较轻、传统唐氏筛查非高风险、NT正常的高龄孕妇。     

三核苷酸重复引物PCR和甲基化特异性多重连接探针扩增技术在脆性X综合征产前诊断中的应用

目的探讨脆性X综合征(FXS)产前基因诊断的方法。方法采用三核苷酸重复引物PCR法(TP-PCR)及甲基化特异性多重连接探针扩增技术(MS-MLPA)检测2个FXS家系及30例正常对照胎儿样本FMR1基因CGG重复数、AGG排布及FMR 1基因Cp G岛甲基化状态。结果正常对照FMR1基因(CGG)n重复数介于23~40之间,频数最大的重复数为29,AGG排布式以9A9A9最为常见。2个FXS家系中前突变等位基因向子代遗传过程中均发生了CGG扩展。正常对照及前突变携带者FMR1基因Cp G岛均为低甲基化状态,全突变患者Cp G岛为高甲基化状态。结论 TP-PCR和MS-PCR联合应用能够检出胎儿FMR1基因CGG重复数和Cp G岛甲基化状态,从而判断胎儿是否为脆性X综合征患儿,为优生干预提供可靠的依据。     

Molecular genetic characterization of a prenatally detected de novo interstitial deletion of chromosome 20p (20p12-p13) encompassing JAG1 and a literature review of prenatal diagnosis of Alagille syndrome

Objective

We present prenatal diagnosis and molecular genetic characterization of a de novo interstitial deletion of chromosome 20p (20p12-p13) and a literature review of prenatal diagnosis of Alagille syndrome (ALGS).

Retrospectively investigating the 12-year experience of prenatal diagnosis of small supernumerary marker chromosomes through array comparative genomic hybridization

Objective

This study retrospectively evaluated the incidences of small supernumerary marker chromosomes (sSMCs) in prenatal diagnoses and detected with gain of pathogenic copy number variation through array comparative genomic hybridization (CGH) in a laboratory in Taiwan.

枫糖尿病患者家系基因突变分析及产前诊断

目的 分析枫糖尿病(maple syrup urine disease,MSUD)患者家系基因突变情况以及对产前诊断的作用. 方法 2005年至2010年间,共诊断4例MSUD患儿,其中男婴和女婴各2例,发病年龄分别为生后2、5、10和26 d.利用聚合酶链反应技术扩增BCKDHA、BCKDHB基因外显子编码区序列,直接测序分析4例患儿BCK DHA、BCKDHB基因的突变情况.在患儿母亲再次妊娠16～20周时抽取羊水细胞,对培养后的羊水细胞进行相应基因突变检测,用于产前诊断.结果 4例患儿中共检测到6种不同的基因突变,包括4种新突变(c.308T＞C、c.562G＞T、c.1279C＞G、c.1280-1291de112).其中在3个家系的BCKDHA基因上检测到5种突变(c.308T＞C、c.562G＞T、c.868G＞A、c.1279C＞G及c.1280-1291de112),另一个家系在BCKDHB基因上检测到c.853C＞T 1个突变.4例患儿母亲再次妊娠羊水细胞中均检测到1个先证者携带的突变,提示胎儿为MSUD杂合子. 结论 通过家系BCKDHA、BCKDHB基因分析,初步了解MSUD患者的基因突变情况,并成功应用于产前诊断,为MSUD家庭提供帮助.     

The application of chromosomal microarray analysis to the prenatal diagnosis of isolated mild ventriculomegaly

Objective

To investigate the clinical value of chromosomal microarray analysis (CMA) in the prenatal diagnosis of genetic abnormalities in fetal isolated mild ventriculomegaly.

Prenatal diagnosis of a familial 5p14.3-p14.1 deletion encompassing CDH18, CDH12, PMCHL1, PRDM9 and CDH10 in a fetus with congenital heart disease on prenatal ultrasound

Objective

We present prenatal diagnosis of a familial 5p14.3-p14.1 deletion in a fetus with congenital heart disease on prenatal ultrasound.

双色共变性荧光原位杂交产前诊断胎儿唐氏综合征

目的 探讨双色共变性荧光原位杂交用于非侵入性产前诊断胎儿唐氏综合征的可行性。方法 对11例孕妇外周血中的胎儿有核红细胞进行抗血型糖蛋白磁珠直接标记,再经磁激活细胞分选法富集,以Y和21号染色体专一探针对分离的胎儿有核红细胞行双色共变性荧光原位杂交,预测胎儿21号染色体倍性和性别,并用羊水染色体核型分析结果,验证预测准确性。结果 11例胎儿21号染色体倍性均正常,与羊水染色体核型分析结果相符。其中5例为男性胎儿,男性胎儿有核红细胞数量为9－65个,平均为25个,男性胎儿有核红细胞纯度为1．4％－18．8％;6例为女性胎儿,孕妇外周血中未见男性胎儿有核红细胞;性别预测结果与羊水染色体型分析结果一致。结论 双色共变性荧光原位杂交用于分析胎儿21号染色体倍性及性别,诊断胎儿唐氏综合征准确、可靠。     

Congenital diaphragmatic hernia, etiology and management, a 10-year analysis of a single center

Objective To analyze congenital diaphragmatic hernia (CDH) during a 10-year period at the University of Kiel, from 1995 through 2004, in order to develop a strategy to improve prenatal diagnosis, to be able to consider endoscopical treatment for selected cases and to assess the current postnatal treatment strategies. Methods Data were obtained from the fetal medicine ultrasound department, from the birth registry, from the postmortem registry, from the neonatal intensive care unit, from pediatric surgery and from the genetic database. Data were subselected for chromosomes, genetic syndromes, for isolated CDH and for associated anomalies, the lung to head ratio and lung volumes were assessed. Data were analyzed respectively for gestation at diagnosis, the type of CDH, the perinatal management and the postnatal outcome. Results There were 29 cases of CDH, in 10/29 (34%) the parents requested termination of pregnancy of which two had already died during pregnancy, 12/19 (63%) survived, which was defined as discharge from the neonatal intensive care unit, seven newborns 7/19 (37%) died in the hospital, 5 of these 5/7 (71%) were delivered in Kiel. A prenatal diagnosis was performed in 16/29 (55%), 1/16 (6%), 7/16 (43%) and 8/16 (50%) in the 1st, 2nd and 3rd trimester, respectively; in 10/29 (34%) diagnosis was performed postpartum, in 3/29 (10%) the diagnosis was performed at autopsy following termination of pregnancy. When the liver was in the abdomen, 9/10 (90%) of the children survived, compared to only 3/8 (43%) when the liver was located in the thorax. A lung to head ratio of 0.81 at 24 weeks resulted in death due to pulmonary hypoplasia. Conclusions The overall survival in CDH is around 50%, antenatal endoscopical therapy may only be considered, if the diagnosis is performed in the early second trimester, and selection criteria such as the lung to head ratio, associated defects and the chromosomal status can be applied.     

先天性畸形的产前诊断、围产期管理及随诊网络的建立与初步实践效果分析

目的 通过先天性畸形的产前诊断、围产期管理和随诊网络的建立,分析初步实践的效果. 方法网络由"产前诊断和分娩中心"与"新生儿治疗及随诊中心"组成.两个中心随时保持业务交流与合作,以实现其网络化功能. 结果网络运行模式:超声科及产科通过超声等影像学检查发现畸形,家长到新生儿外科咨询,新生儿外科与产科、超声科讨论诊断、制定分娩与诊疗计划,按计划分娩,根据病情选择:(1)配有新生儿转运设备的救护车即刻转诊;(2)救护车常规转运;(3)在分娩医院观察及出院后到新生儿外科门诊.转诊患儿进一步检查后根据病情选择急诊手术、择期手术或观察.2003年3月至2007年12月共接诊经产前超声诊断的先天性畸形患儿228例.19例妊娠中期终止妊娠;4例出生后放弃治疗;56例在新生儿期手术,其中51例治愈、5例术后放弃治疗;149例出生后未行手术,仅观察. 结论对先天性畸形的产前诊断、围产期管理及随诊实施网络化管理,进一步加强小儿外科与产科、超声科的密切交流与合作,在实践中逐步建立一套适合我国现实医疗环境、较规范的运作模式,可望进一步提高我国先天性畸形的围产期管理和总体诊治水平.