| 陈宏健,冯桂,王菲.定量荧光聚合酶链式反应在流产组织染色体畸变分析中的诊断价值[J].安徽医药,2024,28(10):2009-2017. |
| 定量荧光聚合酶链式反应在流产组织染色体畸变分析中的诊断价值 |
| The diagnostic value of QF-PCR in chromosomal aberration analysis of abortion tissue |
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| DOI：10.3969/j.issn.1009-6469.2024.10.021 |
| 中文关键词: 染色体畸变 流产，自然 高通量核苷酸序列分析 定量荧光聚合酶链式反应 拷贝数变异测序 流产组织 |
| 英文关键词: Chromosome aberrations Abortion, spontaneous High-throughput nucleotide sequencing QF-PCR CNV-seq Abortion tissue |
| 基金项目:海南省自然科学基金项目( 821RC728)；海南省高等学校科学研究项目( Hnky2019-41)；海南省科技专项( ZDYF2020113) |
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| 中文摘要: |
| 目的评估在以拷贝数变异测序( CNV-seq)为主要遗传学检测手段对流产组织进行染色体畸变分析中辅以定量荧光聚合酶链式反应( QF-PCR)检测的诊断价值。方法采用 QF-PCR及 CNV-seq技术同时检测 2021年 9月至 2022年 11月海口市妇幼保健院收集的 110例流产组织样本。此外， QF-PCR技术将额外检测 50例健康人群外周血样本进行数据补充。 QF-PCR体系为自建体系，在 13、18、21、性染色体共选择 17个位点并将其分为两组。 Panel A组由 D13S796、D13S256、D18S386、D18S535、 D21S11、D21S1446、DXS6809、HPRT构成； Panel B组由 D13S371、D13S634、D18S51、D18S535、D18S819、D21S1409、D21S1412、 DYS218、DXS981、AMEL构成。分析 QF-PCR在判别 13、18、21、性染色体非整倍体时与 CNV-seq结论一致性、 QF-PCR为 CNV-seq进行母源污染鉴定能力，以及 QF-PCR进行 13、18、21、性染色体非整倍体的分型能力。结果对 110例流产组织进行检测，种方法结论一致例数占比为 93.6%。其中 QF-PCR避免了 5例 CNV-seq自身局限性造成的不准确结论， 2例为 QF-PCR的检测失两败。在母源细胞污染鉴定能力上， 15个短串联重复序列的观察杂合度、期望杂合度与个体识别能力值范围分别为 0.737~0.950、0.593~0.941、0.817~0.975；D13、D18、D21与 DX的累积 He依次为 0.999 9、0.999 8、0.999 8与 0.998 5，累积个体识别能力大于0.999999 999 999 999 999 999 999 999 999；自建的 QF-PCR方法在分型能力上表现良好，其干扰因素影响较小。结论 QF-PCR联合 CNV-seq检测可能成为未来孕早期流产组织遗传学分析的主要方法。它不仅能够降低 CNV-seq的误诊率、鉴别母体细胞污染。采用自建 QF-PCR体系还能够显著降低联合检测成本。 |
| 英文摘要: |
| Objective To investigate the assisted diagnostic value of quantitative fluorescent polymerase chain (QF-PCR) in the chromosome aberration analysis of aborted tissue with copy number variation sequencing (CNV-seq) as a main genetic detection method. Methods QF-PCR and CNV-seq were used to detect chromosome aberration in 110 abortion tissue samples which were collectedfrom Haikou Maternal and Child Health Hospital from September 2021 to November 2022. In addition, QF-PCR detected common chro·mosome aberration in an additional 50 blood samples from healthy people for data supplement. QF-PCR system was self-built, consisting of 17 loci selected from 13, 18, 21 and sex chromosomes, respectively. They were divided into two panels. Panel A was comprised ofD13S796, D13S256, D18S386, D18S535, D21S11, D21S1446, DXS6809, HPRT, and Panel B was comprised of D13S371, D13S634,D18S51, D18S535, D18S819, D21S1409, D21S1412, DYS218, DXS981, AMEL. The consistency of the two methods in aneuploid detection of 13, 18, 21 and sex chromosomes, abilities of maternal contamination identification (MCC) and the genotyping of QF-PCR for CNV-seq detection were analyzed.Results A total of 110 cases of abortion tissues were detected, and 93.6% of the cases were consistent with the conclusion of the two methods. Especially, QF-PCR avoided misdiagnosis of CNV-seq in 5 cases, and performed failure in2 cases. In the ability to identify MCC, range of heterozygosity expectation, heterozygosity observation and discrimination power valueof 15 short tandem repeats were 0.737-0.950, 0.593-0.941 and 0.817-0.975, respectively. The cumulative He values of D13, D18, D21and DX were 0.999 9, 0.999 8, 0.999 8 and 0.998 5, respectively. Moreover, CDP was greater than 0.999 999 999 999 999 999 999999 999 999 999. The self-established QF-PCR system in our study displayed favorable genotyping ability, with low-level influence by interference factors.Conclusions QF-PCR combined with CNV-seq may become the main method for genetic analysis of early pregnancy abortion tissue in the future, which can not only reduce the misdiagnosis rate of CNV-seq, but also identify maternal cell contamination. Moreover, a self-established QF-PCR system can significantly reduce the cost of joint detection. |
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