Unit of Hematology for Rare Diseases of Blood and Blood-Forming Organs, Laboratory for Molecular Diagnosis of Rare Hematological Diseases, Azienda Ospedaliera Ospedali Riuniti Villa Sofia Cervello, Palermo, Italy.
Unit of Obstetrical and Gynecology, Fetal Medicine and Prenatal Diagnosis, Azienda Ospedaliera Ospedali Riuniti Villa Sofia Cervello, Palermo, Italy.
J Obstet Gynaecol. 2022 Jul;42(5):1524-1531. doi: 10.1080/01443615.2021.2014429. Epub 2022 Jan 10.
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 Statement 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. 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. 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.
科凯恩综合征 (CS) 是一种罕见的常染色体隐性多系统疾病，其特征是症状早期严重进展。本研究报告了通过在妊娠 8 周时行体腔穿刺术而非羊膜腔穿刺术或绒毛膜取样术进行 CS 产前诊断的可行性。有 3 对 CS 高危夫妇要求通过体腔穿刺术进行产前诊断。在妊娠 8 周的 4 例单胎妊娠中从体腔腔中吸出体腔液，通过微操作器回收 40 个胎儿细胞。通过定量荧光 PCR (QF-PCR) 评估母体 DNA 污染情况，并扩增和测序包含 ERCC6 基因突变的胎儿 DNA 靶区域。在所有这些情况下，均进行了分子分析。1 例胎儿受 CS 影响，流产后胎盘组织证实诊断。在 3 例中，胎儿为父母突变的携带者，分娩后得到证实。本研究表明，在妊娠早期使用存在于体腔液中的胎儿细胞可对 CS 进行可靠的产前诊断。如果已知父母突变，体腔穿刺术可应用于 CS 及其他单基因疾病的极早期妊娠产前诊断。
之前利用体腔穿刺术进行产前胎儿性别鉴定的研究报告的成功率差异很大，范围为 58%至 95%，因为总 DNA 含量低且存在母体细胞污染。这种方法从未在妊娠 8 周时用于罕见的遗传性疾病（如科凯恩综合征）的早期产前诊断。本研究表明，如果已知分子缺陷，体腔液取样结合标准化的实验室程序可应用于妊娠 8 周时的任何罕见的单基因疾病的产前诊断。在对单基因疾病进行体腔穿刺术检查的高危夫妇中进行的这项研究结果表明，从 CF 中选择胚胎-胎儿细胞可实现疾病的可靠和早期产前诊断。这项技术对父母很有吸引力，因为它可以比传统方法至少提前 4 周进行遗传疾病的产前诊断，减轻父母的焦虑，并提供在 8-10 周妊娠时终止受影响病例的选择，这比妊娠中期的手术终止创伤更小、更安全。进一步的研究涉及在妊娠 8 周时获得胎儿核型的可能性和宫内矫正治疗的可能性。
