Abstract：
Cockayne\'s syndrome (CS) is a rare autosomal recessive multisystem disease characterised by early severe progression of symptoms. This study reports the feasibility of earlier prenatal diagnosis of CS by coelocentesis at 8 weeks of gestation respect to amniocentesis or villocentesis. Three couples at risk for CS asked to perform prenatal diagnosis by coelocentesis. Coelomic fluid was aspired from coelomic cavity in four singleton pregnancy at 8 weeks of gestation and 40 foetal cells were recovered by micromanipulator. Maternal DNA contamination was evaluated by quantitative fluorescent PCR (QF-PCR) and target regions of foetal DNA containing parental mutations of ERCC6 gene were amplified and sequenced. In all these cases, molecular analysis was possible. One foetus resulted affected of CS and the diagnosis was confirmed on placental tissue after voluntary abortion. In three cases, foetuses resulted carrier of a parental mutation and the results were confirmed after the birth. This study suggests that reliable prenatal diagnosis of CS could be performed using foetal cells present in coelomatic fluid in earlier pregnancy. Coelocentesis could be applied in prenatal diagnosis of CSs as well as for other monogenic diseases, at very early stage of pregnancy, if parental mutations are already known.Impact StatementWhat is already know on this subject? Previous studies utilising coelocentesis for prenatal determination of foetal sex reported variable success ranging from 58% to 95%, because of low total DNA content and presence of maternal cell contamination. This procedure has never been reported for early prenatal diagnosis at 8 weeks of gestation for rare genetically transmitted diseases such as Cockayne\'s syndrome.What do the results of this study add? This study demonstrates that coelomic fluid sampling combined with well-standardised laboratory procedures can be applied for prenatal diagnosis at eight weeks of gestation for any rare monogenic disease if molecular defects are known.What are the implications of these findings for clinical practice and/or further research? The findings of this study in at risk couples for monogenic diseases investigated by coelocentesis demonstrate that embryo-foetal cell selection from CF allows reliable and early prenatal diagnosis of diseases. This technique is attractive to parents because it provides prenatal diagnosis of genetic disease at least 4 weeks earlier than what can be achieved by the traditional procedures reducing anxiety of parents and provides the option for medical termination of affected cases at 8-10 weeks\' gestation, which is less traumatic and safer than second-trimester surgical termination. Further research concerns the possibility to obtain foetal karyotype at eight weeks of gestation and the possibility of intrauterine corrective therapy.
摘要：
Cockayne综合征（CS）是一种罕见的常染色体隐性多系统疾病，其特征是症状的早期严重进展。这项研究报告了在妊娠8周时通过腔穿刺术对CS进行早期产前诊断的可行性。三对有CS风险的夫妇要求通过腔穿刺术进行产前诊断。在妊娠8周时的四次单胎妊娠中，从体腔中吸取了体腔液，并通过显微操纵器回收了40个胎儿细胞。通过定量荧光PCR（QF-PCR）评估母体DNA污染，并扩增含有ERCC6基因亲本突变的胎儿DNA靶区域并进行测序。在所有这些情况下，分子分析是可能的。一名胎儿受CS影响，自愿流产后在胎盘组织上证实了诊断。在三种情况下，胎儿导致父母突变的携带者，结果在出生后得到证实。这项研究表明，可以在妊娠早期使用存在于腹腔液中的胎儿细胞进行CS的可靠产前诊断。腔穿刺术可用于CSs的产前诊断以及其他单基因疾病。在怀孕的早期阶段，如果亲本突变是已知的。先前的研究利用腔穿刺术进行胎儿性别的产前测定报告的变量成功范围从58%到95%,因为总DNA含量低和母体细胞沾染的存在。这种方法从未在妊娠8周时进行早期产前诊断，例如Cockayne综合征。这项研究的结果增加了什么？这项研究表明，如果已知分子缺陷，则可以在妊娠八周时对任何罕见的单基因疾病进行产前诊断，并结合标准化的实验室程序进行体腔液采样。这些发现对临床实践和/或进一步研究有什么意义？这项研究在腔穿刺术研究的单基因疾病高危夫妇中的发现表明，从CF中选择胚胎胎儿细胞可以可靠和早期地进行疾病的产前诊断。这项技术对父母很有吸引力，因为它提供了遗传病的产前诊断，至少比传统程序提前4周，减少父母的焦虑，并提供了在妊娠8-10周时终止受影响病例的选择。这是较少的创伤和安全比中期妊娠手术终止。进一步的研究涉及在妊娠八周获得胎儿核型的可能性以及宫内矫正治疗的可能性。
