CNVplex联合STR检测技术在自然流产患者流产物中的应用

Application of CNVplex combined with STR detection in products of conception from patients with spontaneous abortion

摘要

目的 评估高通量连接探针技术(CNVplex)结合短串联重复序列(STR)检测技术在分析自然流产患者流产物的遗传学因素中的应用价值。方法 选取2021年1月至2024年12月符合纳入标准的970例自然流产患者为研究对象,采集流产物进行CNVplex及STR检测,分析不同孕期和不同年龄段患者流产物的染色体异常情况。结果 970例流产物中有575例检出染色体数量或结构异常,检出率为59.28%(575/970)。孕早期流产患者流产物共873例,异常染色体检出率60.82%(531/873),其中非整倍体异常(77.97%)和三倍体(16.57%)最常见,结构异常次之(4.52%),嵌合体占0.75%,全基因组单亲二体(UPD)占0.38%;染色体非整倍体中以16三体(24.40%)、22三体(19.57%)、X单体(12.56%)最常见。97例孕中期流产物异常检出率为45.36%(44/97),仅见非整倍体异常(86.36%)和三倍体(13.64%);非整倍体中X单体(34.21%)、16三体(15.79%)、21三体(13.16%)、22三体(7.89%)最常见。孕早期流产物染色体异常总检出率(60.82%,531/873)显著高于孕中期(45.36%,44/97),差异有统计学意义(χ²=8.648,P<0.05)。孕中期流产物中X单体的检出率(13.40%,13/97)显著高于孕早期(5.96%,52/873),差异有统计学意义(χ²=7.741,P<0.05)。801例<35岁患者流产物中染色体异常检出率为56.80%(455/801),169例≥35岁患者流产物染色体异常检出率为71.01%(120/169)。<35岁患者流产物与≥35岁患者相比较,后者染色体异常检出率更高,差异有统计学意义(χ²=11.661,P<0.05)。结论 CNVplex联合STR检测技术有助于分析评估再发风险,为精准的生殖遗传咨询和生育规划指导提供了遗传学依据。

Abstract

Objective To evaluate the application value of high-throughput ligation-dependent probe amplification (CNVplex) combined with short tandem repeat (STR) analysis in identifying genetic factors in products of conception (POC) from patients with spontaneous abortion (SA). Methods A total of 970 patients with spontaneous abortion who met the inclusion criteria between January 2021 and December 2024 were enrolled. Products of conception were collected for CNVplex and STR analyses. Chromosomal abnormalities were analyzed according to gestational stage and maternal age. Results Among the 970 POC samples, chromosomal numerical or structural abnormalities were detected in 575 cases, with a detection rate of 59.28% (575/970). Among 873 cases of early spontaneous abortion, the detection rate of chromosomal abnormalities was 60.82% (531/873). The most common abnormalities were aneuploidy (77.97%) and triploidy (16.57%), followed by structural abnormalities (4.52%), mosaicism (0.75%), and whole-genome uniparental disomy (UPD) (0.38%). Among aneuploidies, trisomy 16 (24.40%), trisomy 22 (19.57%), and monosomy X (12.56%) were the most frequent. Among 97 cases of mid-trimester spontaneous abortion, the overall abnormality detection rate was 45.36% (44/97), including aneuploidy (86.36%) and triploidy (13.64%) only. The most common aneuploidies were monosomy X (34.21%), trisomy 16 (15.79%), trisomy 21 (13.16%), and trisomy 22 (7.89%). The overall chromosomal abnormality detection rate in early spontaneous abortion (60.82%, 531/873) was significantly higher than that in mid-trimester spontaneous abortion (45.36%, 44/97) (χ²=8.648, P<0.05). The detection rate of monosomy X was significantly higher in mid-trimester POC (13.40%, 13/97) than in early POC (5.96%, 52/873) (χ²=7.741, P<0.05). Among 801 patients aged <35 years, the chromosomal abnormality detection rate was 56.80% (455/801), whereas among 169 patients aged ≥35 years, the rate was 71.01% (120/169). Compared with patients <35 years, those ≥35 years had a significantly higher detection rate of chromosomal abnormalities (χ²=11.661, P<0.05). Conclusion The combination of CNVplex and STR detection is useful for analyzing chromosomal abnormalities in POC from patients with spontaneous abortion, facilitating evaluation of recurrence risk and providing a genetic basis for precise reproductive genetic counseling and fertility planning.

关键词

高通量连接探针技术 / 短串联重复序列 / 自然流产 / 流产物 / 染色体异常

Key words

CNVplex / short tandem repeat / spontaneou abortion / apoblema / chromosomal abnormality

参考文献

[1] Quenby S, Gallos I D, Dhillon-Smith R K, et al. Miscarriage matters: the epidemiological, physical, psychological, and economic costs of early pregnancy loss[J]. Lancet, 2021, 397(10285): 1658-1667.

[2] La X, Wang W, Zhang M, et al. Definition and multiple factors of recurrent spontaneous abortion[J]. Adv Exp Med Biol, 2021, 1300: 231-257.

[3] Dai Y F, Wu X Q, Huang H L, et al. Experience of copy number variation sequencing applied in spontaneous abortion[J]. BMC Med Genomics, 2024, 18(1): 45.

[4] Melo P, Dhillon-Smith R, Islam M A, et al. Genetic causes of sporadic and recurrent miscarriage[J]. Fertil Steril, 2023, 120(5): 940-944.

[5] Dimitriadis E, Menkhorst E, Saito S, et al. Recurrent pregnancy loss[J]. Nat Rev Dis Primers, 2020, 6(1): 98.

[6] 朱思静, 甄帅, 牛惠, 等. 产前诊断领域全外显子组测序的应用现况与未来展望[J]. 中国妇幼健康研究, 2025, 36(9): 61-67.

[7] Xu J, Hu L, Wen L, et al. Detection of a novel large fragment deletion in the alpha-globin gene cluster using the CNVplex technology[J]. Front Genet, 2025, 16: 1518392.

[8] 中华预防医学会出生缺陷预防与控制专业委员会, 中国优生科学协会基因诊断与精准医学分会, 胡婷, 等. 拷贝数变异检测在产前诊断中的应用指南[J]. 中华医学遗传学杂志, 2020, 37(9): 909-917.

[9] 袁思敏, 徐霞, 廖灿, 等. 335例早期自然流产患者胚胎绒毛染色体分析[J]. 中国妇幼健康研究, 2020, 31(9): 1190-1198.

[10] 侯东霞, 侯丽青, 乌云塔娜, 等. 基于微阵列技术分析胚胎停育绒毛组织中的染色体异常[J]. 实用医学杂志, 2020, 36(19): 2655-2660.

[11] Chen L, Wang L, Tang F, et al. Copy number variation sequencing combined with quantitative fluorescence polymerase chain reaction in clinical application of pregnancy loss[J]. J Assist Reprod Genet, 2021, 38(9): 2397-2404.

[12] Luo S, Chen X, Yan T, et al. Application of copy number variation sequencing in genetic analysis of miscarriages in early and middle pregnancy[J]. Cytogenet Genome Res, 2020, 160(11-12): 634-642.

[13] 陈闫, 刘伟, 兰智辉, 等. CNV-seq联合STR在胚胎停滞发育患者妊娠产物中的应用[J]. 检验医学与临床, 2024, 21(23): 3572-3575.

[14] 张元宝, 徐晓莲, 黄存, 等. 拷贝数变异测序结合短串联重复序列分析应用于流产物分析及产前诊断[J]. 中华医学遗传学杂志, 2021, 38(6): 577-580.

[15] 黄霈, 陈美佳, 吕福通, 等. 早期自然流产胚胎绒毛染色体多重连接依赖式探针扩增技术分析[J]. 中国计划生育和妇产科, 2020, 12(5): 39-42.

[16] 夏政怡, 周冉, 孟露露, 等. 染色体微阵列分析技术在偶发自然流产遗传学诊断中的应用[J]. 现代妇产科进展, 2022, 31(3): 191-195.

[17] 胡柳琴, 黄燕飞, 陈敏, 等. 产前超声联合胎盘病理在三倍体综合征中的诊断价值[J]. 中国产前诊断杂志(电子版), 2024, 16(4): 28-34.

[18] 罗艳, 尹恒. 高龄产妇妊娠并发症及妊娠结局影响因素分析[J]. 中国妇幼健康研究, 2021, 32(11): 1595-1599.

[19] Jiang X, Yan J, Sheng Y, et al. Low anti-Mullerian hormone concentration is associated with increased risk of embryonic aneuploidy in women of advanced age[J]. Reprod Biomed Online, 2018, 37(2): 178-183.

[20] 罗珊, 李小红, 汪燕, 等. 胚胎植入前非整倍体遗传学检测对高龄女性活产率的影响及相关因素分析[J]. 四川大学学报(医学版), 2024, 55(5): 1288-1294.

[21] Bai W, Zhang Q, Lin Z, et al. Analysis of copy number variations and possible candidate genes in spontaneous abortion by copy number variation sequencing[J]. Front Endocrinol (Lausanne), 2023, 14: 1218793.

[22] Correa-De-Araujo R, Yoon S. Clinical outcomes in high-risk pregnancies due to advanced maternal age[J]. J Womens Health (Larchmt), 2021, 30(2): 160-167.

基金资助

成都医学院临床科学研究基金项目(24LHFYSZ1-21)