中华检验医学杂志
中華檢驗醫學雜誌
중화검험의학잡지
CHINESE JOURNAL OF LABORATORY MEDICINE
2011年
3期
236-239
,共4页
王明山%金艳慧%牛真珍%谢海啸%谢耀盛%杨丽红
王明山%金豔慧%牛真珍%謝海嘯%謝耀盛%楊麗紅
왕명산%금염혜%우진진%사해소%사요성%양려홍
因子Ⅶ缺乏症%血液凝集障碍,遗传性%突变%聚合酶链反应
因子Ⅶ缺乏癥%血液凝集障礙,遺傳性%突變%聚閤酶鏈反應
인자Ⅶ결핍증%혈액응집장애,유전성%돌변%취합매련반응
Factor Ⅶ deficiency%Blood coagulation disorders,inherited%Mutation%Polymerase chain reaction
目的 对一个遗传性FⅦ缺乏症家系进行F7基因突变检测,探讨其分子发病机制.方法 采用ELISA法检测先证者及家系成员FⅦ∶Ag,采用一期凝固法检测先证者及家系成员PT、FⅦ∶C等凝血指标进行实验表型诊断.基因检测采用DNA直接测序法分析先证者及家系成员F7基因的全部外显子及侧翼、5'和3'非翻译区,发现突变位点用反向测序加以证实;用CLC Protein Workbench软件分析基因突变位点的物种保守性和蛋白质二级结构改变.选择100名健康对照者以排除基因多态性.结果 先证者及其妹妹的PT、FⅦ∶C、FⅦ∶Ag明显异常,分别为36.3 s、5.0%、40.7%和33.4 s、5.0%、37.4%;先证者的父亲、母亲、女儿、外甥女的PT稍延长,分别为14.9 s、14.6 s、15.5 s、14.6 s;其FⅦ:C稍减低,分别为70%、85%、59%、79%.先证者F7基因8号外显子c.784T＞C杂合突变导致Ser269Pro,8号外显子c.964T＞G杂合突变导致Cys329Gly;先证者妹妹为c.784T＞C和c.964T＞G双重杂合突变,母亲为c.784T＞C杂合子,父亲、女儿、外甥女均为c.964T＞G杂合子.蛋白质生物学特性分析发现,Cys329Gly导致蛋白质空间构型发生改变,Ser269Pro导致氨基酸极性及疏水性发生改变.结论 F7基因Ser269Pro和Cys329Gly双重杂合突变是导致该例遗传性FⅦ缺乏症的分子发病机制.
目的 對一箇遺傳性FⅦ缺乏癥傢繫進行F7基因突變檢測,探討其分子髮病機製.方法 採用ELISA法檢測先證者及傢繫成員FⅦ∶Ag,採用一期凝固法檢測先證者及傢繫成員PT、FⅦ∶C等凝血指標進行實驗錶型診斷.基因檢測採用DNA直接測序法分析先證者及傢繫成員F7基因的全部外顯子及側翼、5'和3'非翻譯區,髮現突變位點用反嚮測序加以證實;用CLC Protein Workbench軟件分析基因突變位點的物種保守性和蛋白質二級結構改變.選擇100名健康對照者以排除基因多態性.結果 先證者及其妹妹的PT、FⅦ∶C、FⅦ∶Ag明顯異常,分彆為36.3 s、5.0%、40.7%和33.4 s、5.0%、37.4%;先證者的父親、母親、女兒、外甥女的PT稍延長,分彆為14.9 s、14.6 s、15.5 s、14.6 s;其FⅦ:C稍減低,分彆為70%、85%、59%、79%.先證者F7基因8號外顯子c.784T＞C雜閤突變導緻Ser269Pro,8號外顯子c.964T＞G雜閤突變導緻Cys329Gly;先證者妹妹為c.784T＞C和c.964T＞G雙重雜閤突變,母親為c.784T＞C雜閤子,父親、女兒、外甥女均為c.964T＞G雜閤子.蛋白質生物學特性分析髮現,Cys329Gly導緻蛋白質空間構型髮生改變,Ser269Pro導緻氨基痠極性及疏水性髮生改變.結論 F7基因Ser269Pro和Cys329Gly雙重雜閤突變是導緻該例遺傳性FⅦ缺乏癥的分子髮病機製.
목적 대일개유전성FⅦ결핍증가계진행F7기인돌변검측,탐토기분자발병궤제.방법 채용ELISA법검측선증자급가계성원FⅦ∶Ag,채용일기응고법검측선증자급가계성원PT、FⅦ∶C등응혈지표진행실험표형진단.기인검측채용DNA직접측서법분석선증자급가계성원F7기인적전부외현자급측익、5'화3'비번역구,발현돌변위점용반향측서가이증실;용CLC Protein Workbench연건분석기인돌변위점적물충보수성화단백질이급결구개변.선택100명건강대조자이배제기인다태성.결과 선증자급기매매적PT、FⅦ∶C、FⅦ∶Ag명현이상,분별위36.3 s、5.0%、40.7%화33.4 s、5.0%、37.4%;선증자적부친、모친、녀인、외생녀적PT초연장,분별위14.9 s、14.6 s、15.5 s、14.6 s;기FⅦ:C초감저,분별위70%、85%、59%、79%.선증자F7기인8호외현자c.784T＞C잡합돌변도치Ser269Pro,8호외현자c.964T＞G잡합돌변도치Cys329Gly;선증자매매위c.784T＞C화c.964T＞G쌍중잡합돌변,모친위c.784T＞C잡합자,부친、녀인、외생녀균위c.964T＞G잡합자.단백질생물학특성분석발현,Cys329Gly도치단백질공간구형발생개변,Ser269Pro도치안기산겁성급소수성발생개변.결론 F7기인Ser269Pro화Cys329Gly쌍중잡합돌변시도치해례유전성FⅦ결핍증적분자발병궤제.
Objective To identify gene mutations and explore the molecular mechanism of a pedigree with inherited coagulation F Ⅶ deficiency. Methods The levels of F Ⅶ: Ag in the proband and other family members were measured by ELISA assay. The values of PT, F Ⅶ: C and other coagulant parameters were determined by one-stage clotting for laboratory phenotype diagnosis. All the exons,exon-intron boundaries and 5',3' untranslated sequences of F7 gene were amplified by direct sequencing. The detected mutations were further confirmed by sequencing the other stand. The CLC Protein Workbench software was used to analyze the species conservation of the mutated site and the protein secondary structure. 100 healthy individuals were selected to exclude gene polymorphism. Results PT, FⅦ∶C and FⅦ: Ag in the proband and his sister were abnormal, which were 36. 3 s, 5.0%, 40. 7% and 33.4 s,5. 0%, 37.4%, respectively. Both PT and FⅦ∶C in the proband's father, mother, daughter and niece were slightly abnormal, which were 14.9 s, 14. 6 s, 15.5 s, 14. 6 s and 70%, 85%, 59%, 79%, respectively.The heterozygous mutations c. 784T ＞ C and c. 964T ＞ G in exon 8 of F7 gene were found in the proband,resulting in the substitutions of Ser269Pro and Cys329Gly respectively. Compound heterozygous mutations c. 784T ＞ C and c. 964T ＞ G were found in the proband's sister. The proband's mother was heterozygous for c. 784T ＞ C. His father, daughter and niece were heterozygous for c. 964T ＞ G. The protein biological characteristics analysis revealed that the Cys329Gly caused the change of spatial configuration, and Ser269Pro led to the change of amino acid polarity and hydrophobicity. Conclusion Compound heterozygous mutations of Cys329Gly and Ser269 Pro in F7 gene may be the underlying molecule mechanism of FⅦ deficiency in this pedigree.
