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,两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0.9999、0.9998、0.9998与0.9985,累积个体识别能力大于0.999999999999999999999999999999;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。
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 collected from Haikou Maternal and Child Health Hospital from September 2021 to November 2022.In addition,QF-PCR detected common chromosome 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 of D13S796,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 in 2 cases.In the ability to identify MCC,range of heterozygosity expectation,heterozygosity observation and discrimination power value of 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,D21 and DX were 0.9999,0.9998,0.9998 and 0.9985,respectively.Moreover,CDP was greater than 0.999999999999999999999999999999999.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 ra
作者
陈宏健
冯桂
王菲
CHEN Hongjian;FENG Gui;WANG Fei(Department of Reproductive Medicine,The First Affiliated Hospital of Hainan Medical University,Haikou,Hainan 570102,China;Medical Genetics and Prenatal Diagnosis,Haikou Maternal and Child Health Hospital,Haikou,Hainan 570203,China)
出处
《安徽医药》
CAS
2024年第10期2009-2017,共9页
Anhui Medical and Pharmaceutical Journal
基金
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)。
