Application of NICS Testing at Different Stages in PGT Treatment of Patients with Reciprocal Translocation
-
摘要:
目的 评估不同时期NICS检测在平衡易位患者PGT治疗中应用的可行性。 方法 分析检测结果（包括检测成功和检测失败）和CNVs结果（包括正常和和异常）2个变量；根据样本类别，分成D3培养液（D3 NICS）、D5培养液（D5 NICS）和整胚及活检细胞（胚胎/TE）组，对各组之间的检测失败率和CNVs结果分布进行比较。 结果 D3 NICS检测失败率和CNVs结果分布同胚胎/TE组相比，差异有统计学意义（P < 0.001/P < 0.01）；D5 NICS组的检测失败率结果和CNVs结果分布同胚胎/TE组相比，差异无统计学意义（P > 0.05）。 结论 相对于D3 NICS，D5 NICS在平衡易位患者胚胎的诊断应用上更有优势。作为一种无损检测手段，随着操作方法和测序技术的进步，有望替代传统活检技术用于平衡易位患者胚胎的临床诊断。 Abstract:Objective To study the application of NICS of different stages in the PGT treatment for patients with balanced translocation. Methods The parameters analyzed included testing results （tested successfully and rested failed） and CNVs results （normal and abnormal）; the samples were divided into D3 NICS, D5 NICS, whole embryos and biopsied cells （embryo /TE） groups according to the samples types. The testing failure rate and CNVs results distribution were compared among the groups. Results There were significant differences between D3 NICS group and embryo/TE group on testing failure rate and CNVs results distribution （P < 0.001/P < 0.01）, while no significant difference was shown in results between D5 NICS group and embryo /TE group （P > 0.05）. Conclusions Compared with D3 NICS, D5 NICS has more advantages in the application of balanced translocation embryos diagnosis. As a non-invasive test, D5 NICS is expected to replace the traditional biopsy technique in treatment of balanced translocation patients with the progress of manipulation methods and sequencing technology. -
Key words:
- Reciprocal translocation /
- NICS /
- PGT
-
表 1 各组样本CNV检出结果汇总[n（%）]
Table 1. Summary of CNV results in different groups [n（%）]
变量 合计 D3 NICS D5 NICS 胚胎/TE χ2 P 检测结果 28.805 < 0.001* 检测失败（N/A） 25（13.44） 15（38.46）* 8（11.11） 2（2.67） 检测成功（非N/A） 161（86.56） 24（61.54）* 64（88.89） 73（97.33） CNVs结果 9.806 0.01* 正常 50（31.06） 14（58.33）* 17（26.56） 19（26.03） 异常 111（68.94） 10（41.67）* 47（73.44） 54（73.97） *P < 0.05。 
下载: 导出CSV
表 2 相同胚胎D5 NICS和胚胎/TE 测序结果比较[n（%）]
Table 2. Comparison of sequencing results between D5 NICS group and embryo/TE from the same embryo [n（%）]
变量 合计 D5 NICS 胚胎/TE χ2 P 检测结果 2.972 0.085 检测失败（N/A） 9（6.52） 7（10.14） 2（2.90） 检测成功（非N/A） 129（93.48） 62（89.86） 67（97.10） CNVs结果 0.005* 0.944 正常 35（27.13） 17（27.42） 18（26.87） 异常 94（72.87） 45（72.58） 49（73.13） *P < 0.05。
下载: 导出CSV
表 3 相同胚胎D3 NICS和胚胎/TE测序结果比较[n（%）]
Table 3. Comparison of sequencing results between D3 NICS group and embryo/TE from the same embryo [n（%）]
变量 合计 D3 NICS 胚胎/TE χ2 P 检测结果 15.750 < 0.001* 检测失败（N/A） 16（22.22） 15（41.67） 1（2.78） 检测成功（非N/A） 56（77.78） 21（58.33） 35（97.22） CNVs结果 10.039 < 0.01* 正常 20（35.71） 13（61.90） 7（20.00） 异常 36（64.29） 8（38.10） 28（80.00） *P < 0.05。
下载: 导出CSV
表 4 不同阶段 NICS同胚胎/TE测序结果比较[n（%）]
Table 4. Comparison of sequencing results in NICS groups of different stages and embryo/TE group [n（%）]
变量 合计 D3 NICS D5 NICS 胚胎/TE χ2 P 检测结果 12.917 < 0.01* 检测失败（N/A） 18（20.00） 12（40.00）* 5（16.67） 1（3.33） 检测成功（非N/A） 72（80.00） 18（60.00）* 25（83.33） 29（96.67） CNVs结果 11.997 < 0.01* 正常 21（29.17） 11（61.11）* 4（16.00） 6（20.69） 异常 51（70.83） 7（38.89）* 21（84.00） 23（79.31） *P < 0.05。
下载: 导出CSV
-
[1] Jacobs P A,Browne C,Gregson N,et al. Estimates of the frequency of chromosome abnormalities detectable in unselected newborns using moderate levels of banding[J]. J Med Genet,1992,29(2):103-108. doi: 10.1136/jmg.29.2.103 [2] Ozawa N,Maruyama T,Nagashima T,et al. Pregnancy outcomes of reciprocal translocation carriers who have a history of repeated pregnancy loss[J]. Fertil Steril,2008,90(4):1301-1304. doi: 10.1016/j.fertnstert.2007.09.051 [3] 江美燕,偶健,李红. 染色体平衡易位携带者生育风险评估[J]. 南京医科大学学报,2015,35(9):1291-1296. [4] 郭彩琴,王峻峰,王俊,等. 69例染色体平衡易位携带者的产前诊断与遗传咨询[J]. 中国妇幼保健,2015,30(32):5602-5606. [5] Minasi M G,Colasante A,Riccio T,et al. Correlation between aneuploidy,standard morphology evaluation and morphokinetic development in 1730 biopsied blastocysts: a consecutive case series study[J]. Hum Reprod,2016,31(10):2245-2254. doi: 10.1093/humrep/dew183 [6] Fesahat,F,Montazeri F,Hoseini S M. Preimplantation genetic testing in assisted reproduction technology[J]. J Gynecol Obstet Hum Reprod,2020,49(5):101723. doi: 10.1016/j.jogoh.2020.101723 [7] Xu J,Fang R,Chen L,et al. Noninvasive chromosome screening of human embryos by genome sequencing of embryo culture medium for in vitro fertilization[J]. Proc Natl Acad Sci U S A,2016,113(42):11907-11912. doi: 10.1073/pnas.1613294113 [8] Rubio C,Rodrigo L,Garcia-Pascual C,et al. Clinical application of embryo aneuploidy testing by next-generation sequencing[J]. Biol Reprod,2019,101(6):1083-1090. doi: 10.1093/biolre/ioz019 [9] Russell S J,Menezes K,Balakier H,et al. Comprehensive profiling of Small RNAs in human embryo-conditioned culture media by improved sequencing and quantitative PCR methods[J]. Syst Biol Reprod Med,2020,66(2):129-139. doi: 10.1080/19396368.2020.1716108 [10] Sialakouma A,Karakasiliotis I,Ntala V,et al. Embryonic Cell-free DNA in Spent Culture Medium: A Non-invasive Tool for Aneuploidy Screening of the Corresponding Embryos[J]. In Vivo,2021,35(6):3449-3457. doi: 10.21873/invivo.12645 [11] Rubio C,Navarro-Sánchez L,García-Pascual C M,et al. Multicenter prospective study of concordance between embryonic cell-free DNA and trophectoderm biopsies from 1301 human blastocysts[J]. Am J Obstet Gynecol,2020,223(5):1-13. doi: 10.1016/j.ajog.2020.04.035 [12] Chen L,Sun Q,Xu J,et al. A Non-invasive Chromosome Screening Strategy for Prioritizing in vitro Fertilization Embryos for Implantation[J]. Front Cell Dev Biol,2021,9:708322. doi: 10.3389/fcell.2021.708322 [13] Galluzzi L,Palini S,De Stefani S,et al. Extracellular embryo genomic DNA and its potential for genotyping applications[J]. Future Sci OA,2015,1(14):FSO62. [14] Hammond E R, Shelling A N, Cree L M, Nuclear and mitochondrial DNA in blastocoele fluid and embryo culture medium: evidence and potential clinical use[J]. Hum Reprod, 2016, 31（8）: 1653-1661. [15] Vera-Rodriguez M,Rubio C. Assessing the true incidence of mosaicism in preimplantation embryos[J]. Fertil Steril,2017,107(5):1107-1112. doi: 10.1016/j.fertnstert.2017.03.019 [16] Popovic M,Dhaenens L,Taelman J,et al. Extended in vitro culture of human embryos demonstrates the complex nature of diagnosing chromosomal mosaicism from a single trophectoderm biopsy[J]. Hum Reprod,2019,34(4):758-769. doi: 10.1093/humrep/dez012 [17] Popovic,M,Dheedene A,Christodoulou C,et al. Chromosomal mosaicism in human blastocysts: the ultimate challenge of preimplantation genetic testing?[J]. Hum Reprod,2018,33(7):1342-1354. doi: 10.1093/humrep/dey106 [18] Shitara A,Takahashi K,Goto M,et al. Cell-free DNA in spent culture medium effectively reflects the chromosomal status of embryos following culturing beyond implantation compared to trophectoderm biopsy[J]. PLoS One,2021,16(2):e0246438. doi: 10.1371/journal.pone.0246438 -
