妊娠合并抗凝血酶缺陷症的基因检测和分析

doi:10.3969/j.issn.1673-8640.2024.02.006

摘要/Abstract

摘要：

目的对1例妊娠合并遗传性抗凝血酶(AT)缺陷症患者进行基因检测和分析。方法收集患者的临床资料，检测其血浆抗凝血酶活性(AT：A)、凝血酶原时间(PT)、活化部分凝血活酶时间(APTT)、凝血酶时间(TT)、纤维蛋白原(Fib)、D-二聚体(DD)、纤维蛋白原降解产物(FDP)、蛋白C活性(PC：A)、蛋白S活性(PS：A)等凝血指标。对患者的PROC、PROS1、SERPINC1、SERPIND1、PLG、PROCR、THBD、ADAMTS13、HRG、TFPI、CPB2、HABP2、PLAT、PLAU、SERPINA10、SERPINE1、SERPINF2、CALR、GP6、JAK2、MPL和凝血因子相关基因的转录起始点、5'-非翻译区(UTR)、编码区、剪切点8 bp内含子序列区域、转录终止子点突变、小缺失/重复和剪切位点进行突变检测。采用Sanger测序验证变异位点。对变异位点进行生物信息学分析，以明确致病性。结果患者AT：A降低。基因检测结果显示，患者SERPINC1基因第2号外显子发生c.380G>A(p.Cys127Tyr)杂合突变，JAK2基因第10号外显子发生c.1324G>C(p.Glu442Gln)杂合突变，SERPINA10基因第2号外显子发生c.389T>G(p.Leu130Trp)杂合突变。SERPINC1基因c.380G>A(p.Cys127Tyr)变异在正常人群公共SNP数据库(ExAC数据库、1000G dbSNP13数据库和gnomAD数据库)中未被收录，ClinVar数据库、OMIM数据库和HGMD数据库中均未见该变异位点的相关报道，PolyPhen-2、MutationTaster和CADD在线软件预测其为致病性变异。JAK2基因c.1324G>C(p.Glu442Gln)变异和SERPINA10基因c.389T>G(p.Leu130Trp)评级均为意义不明确的变异。Sanger测序结果显示3个变异均存在。蛋白模拟结构分析结果显示，SERPINC1基因c.380G>A(p.Cys127Tyr)变异可导致蛋白结构改变，从而影响蛋白功能。结论 SERPINC1基因c.380G>A(p.Cys127Tyr)变异可能导致遗传性AT缺陷症。监测凝血相关指标，及时进行分子诊断，有助于改善遗传性AT患者的妊娠结局。

关键词: SERPINC1基因, 易栓症, 遗传性抗凝血酶缺陷症, 妊娠

Abstract:

Objective To analyze and perform genetic testing for a case of pregnancy with antithrombin deficiency. Methods The clinical data were collected，and plasma antithrombin activity(AT：A)，prothrombin time(PT)，activated partial thromboplastin time(APTT)，thrombin time(TT)，fibrinogen(Fib)，D-dimer(DD)，fibrinogen degradation product(FDP)，protein C activity(PC：A) and protein S activity(PS：A)were determined. The proband genes(PROC，PROS1，SERPINC1，SERPIND1，PLG，PROCR，THBD，ADAMTS13，HRG，TFPI，CPB2，HABP2，PLAT，PLAU，SERPINA10，SERPINE1，SERPINF2，CALR，GP6，JAK2，MPL)，coagulation factor related gene at the transcription start point，5'-untranslated region(UTR)，coding region，splicing point 8 bp intron sequence region，transcription terminator point mutation，small deletion or duplication and splicing site were determined for the presence of mutations. Sanger sequencing was used to validate the mutation sites. Bioinformatics analysis of the mutation sites was performed to clarify pathogenicity. Results The patient had decreased AT：A. The patient had a c.380G>A(p.Cys127Tyr)heterozygous mutation in exon 2 of the SERPINC1 gene，a c.1324G>C(p.Glu442Gln) heterozygous mutation in exon 10 of the JAK2 gene，and a c.389T>G(p.Leu130Trp)heterozygous mutation in exon 2 of the SERPINA10 gene. The c.380G>A(p.Cys127Tyr) mutation in the SERPINC1 gene was not included in the public SNP databases for normal populations(ExAC database，1000G dbSNP13 database and gnomAD)，and no relevant reports of this site were seen in the ClinVar database，OMIM database and HGMD database. PolyPhen-2，MutationTaster and CADD online software predicted it as a pathogenic mutation. The c.1324G>C(p.Glu442Gln) mutation in the JAK2 gene and the c.389T>G(p.Leu130Trp) mutation in the SERPINA10 gene were both classified as mutations of uncertain significance. Sanger sequencing results showed the presence of all 3 mutations. Structural analysis of protein mimicry showed that the c.380G>A(p.Cys127Tyr) mutation in the SERPINC1 gene can lead to structural changes in the protein，thereby affecting protein function. Conclusions The c.380G>A(p.Cys127Tyr)mutation in the SERPINC1 gene may lead to hereditary AT deficiency. The monitoring of coagulation-related indicators and timely molecular diagnosis can help improve pregnancy outcomes in patients with hereditary AT.

Key words: SERPINC1 gene, Thrombosis, Heredity antithrombin deficiency, Pregnancy

中图分类号:

R446.7

王怡, 吴瑛婷, 陈慧芬, 李国华, 鲍时华, 戴菁, 王学锋. 妊娠合并抗凝血酶缺陷症的基因检测和分析[J]. 检验医学, 2024, 39(2): 132-137.

WANG Yi, WU Yingting, CHEN Huifen, LI Guohua, BAO Shihua, DAI Jing, WANG Xuefeng. Genetic testing and analysis of pregnancy complicated with antithrombin deficiency[J]. Laboratory Medicine, 2024, 39(2): 132-137.

图/表 4

表1 本例患者凝血指标检测结果

项目	检测值	参考区间
PC：A /%	92.3	70.0~140.0
PS：A/%	95.3	60.0~130.0
AT：A/%	50.0	75.0~125.0
PAI/(U·mL^-1)	3.19	0.30~3.50
PT/s	11.0	9.1~15.1
APTT/s	27.9	18.4~38.4
TT/s	18.9	14.0~21.0
Fib/(g·L^-1)	2.1	2.0~5.0
DD(mg·L^-1)	0.85	<0.55
FDP/(μg·mL^-1)	3.6	<5.0
INR	0.9	0.8~1.1

图1 本例患者基因检测结果注：(a)SERPINC1基因c.380G>A(p.Cys127Tyr)；(b)JAK2基因c.1324G>C(p.Glu442Gln)；(c)SERPINA10基因c.389T>G(p.Leu130Trp)。

图2 变异位点氨基酸序列保守性分析

图3 蛋白结构预测分析注：(a)野生型蛋白；(b)突变型蛋白；箭头所指(红色标记)为127位氨基酸的空间位置。

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