Kahraman S, Benkhalifa M, Donmez E, Biricik A, Sertyel S, Findikli N, Berkil H
Istanbul Memorial Hospital, ART and Genetics Unit, Istanbul, Turkey.
Prenat Diagn. 2004 Apr;24(4):307-11. doi: 10.1002/pd.842.
Preimplantation genetic diagnosis for aneuploidy screening (PGD-AS) using sequential in situ hybridization was applied for aneuploidy testing in 276 couples with 282 ART cycles. Patients with advanced maternal age (AMA, n = 147), recurrent implantation failure (RIF, n = 48), repeated early spontaneous abortion (RSA, n = 32) and abnormal gamete cell morphology (AGCM, n = 55) including macrocephal sperm forms or cytoplasmic granular oocytes were included. Embryo biopsy was performed on day 3 in a calcium-magnesium-free medium by using a noncontact diode laser system. After fixation and enzymatic treatment, fluorescent in situ hybridization (FISH) was carried out on 1147 blastomeres with specific probes for chromosomes 13, 16, 18, 21 and 22 for AMA group, 13, 18, 21, X and Y for AGCM group and 13, 16, 18, 21, 22, X and Y for RIF and RSA groups respectively. The overall chromosomal abnormality rate in analyzed embryos was 40.9%, with no significant difference between AMA, RIF and RSA groups (p > 0.05). However, AGCM group presented a higher rate of chromosomal aneuploidies (57.4%) than the other three groups (p < 0.01). A total of 84% biopsied embryos presented cleavage in 24 h and embryo transfer was realized in 278 cycles. In four cycles, no chromosomally normal embryo was found for embryo transfer. A total of 88 pregnancies (31.6%) were achieved, 19.3% resulted in abortion and 63 healthy births were obtained, with a total of 93 babies born. Aneuploidy testing in couples with poor prognosis undergoing ART cycles is a useful tool to increase the chance of ART success. Furthermore, abnormal gamete cell morphology should be considered one of the major indications for PGD in ART programs as high aneuploidy rates were observed in this group.
采用序贯原位杂交技术进行植入前非整倍体筛查（PGD-AS），对276对夫妇的282个辅助生殖周期进行非整倍体检测。纳入的患者包括高龄产妇（AMA，n = 147）、反复种植失败（RIF，n = 48）、反复早期自然流产（RSA，n = 32）以及配子细胞形态异常（AGCM，n = 55），后者包括大头精子形态或细胞质颗粒状卵母细胞。在无钙镁培养基中于第3天使用非接触式二极管激光系统进行胚胎活检。固定和酶处理后，分别对AMA组的1147个卵裂球用针对13、16、18、21和22号染色体的特异性探针进行荧光原位杂交（FISH），AGCM组用针对13、18、21、X和Y染色体的探针，RIF组和RSA组用针对13、16、18、21、22、X和Y染色体的探针。分析的胚胎中总体染色体异常率为40.9%，AMA组、RIF组和RSA组之间无显著差异（p > 0.05）。然而，AGCM组的染色体非整倍体率（57.4%）高于其他三组（p < 0.01）。总共84%的活检胚胎在24小时内出现卵裂，278个周期实现了胚胎移植。在4个周期中，未发现染色体正常的胚胎用于移植。总共获得88例妊娠（31.6%），19.3%发生流产，获得63例健康分娩，共出生93名婴儿。对预后不良的接受辅助生殖周期的夫妇进行非整倍体检测是增加辅助生殖成功率的有用工具。此外，由于该组观察到高非整倍体率，配子细胞形态异常应被视为辅助生殖项目中PGD的主要指征之一。
