中华血液学杂志
中華血液學雜誌
중화혈액학잡지
Chinese Journal of Hematology
2011年
2期
99-102
,共4页
秦欢欢%王学锋%丁秋兰%许冠群%张利伟%戴菁%陆晔玲%奚晓东%王鸿利
秦歡歡%王學鋒%丁鞦蘭%許冠群%張利偉%戴菁%陸曄玲%奚曉東%王鴻利
진환환%왕학봉%정추란%허관군%장리위%대정%륙엽령%해효동%왕홍리
血管性血友病因子%血管性血友病%DNA突变分析
血管性血友病因子%血管性血友病%DNA突變分析
혈관성혈우병인자%혈관성혈우병%DNA돌변분석
Von Willebrand's factor%Von Willebrand disease%DNA mutational analysis
目的 对3个遗传性血管性血友病(vWD)家系进行表型诊断和基因型分析,探讨其分子发病机制.方法 分别采用出血时间(BT)、活化部分凝血活酶时间(APTT)、瑞斯托霉素诱导的血小板凝集试验(RIPA)、血管性血友病因子(vWF)瑞斯托霉素辅因子(vWF∶RCo)、vWF抗原(vWF∶Ag)、vWF活性(vWF∶A)测定,vWF胶原结合试验(vWF∶CB),vWF与FⅧ结合试验(vWF∶FⅧ∶B)和多聚体分析对3例vWD先证者和家系成员进行表型诊断.提取3例先证者外周血基因组DNA,用PCR法扩增vWF基因的52个外显子及侧翼序列,通过直接测序分析vWF基因变异.结果 3例左证者APTT均延长,BT除先证者3外均正常,血浆RIPA、vWF∶RCo、vWF∶Ag、vWF∶A和vWF∶CB均有不同程度的减低.多聚体分析结果显示先证者3中、高分子质量多聚体缺失,而先证者l、2基本正常.对3例先证者进行基因测序,共发现3个杂合突变,分别为vWF基因的G144067A(R2287Q)、G110374A(R1374H)、C110770T(S1506L)以及28号外显子G110667A(D1472H)杂合多态性.结论 R2287Q、R1374H和S1506L这3种杂合错义突变及D1472H杂合多态性是分别导致3例先证者发生遗传性vWD的分子发病机制.其中R2287Q是国际首次报道的新突变.
目的 對3箇遺傳性血管性血友病(vWD)傢繫進行錶型診斷和基因型分析,探討其分子髮病機製.方法 分彆採用齣血時間(BT)、活化部分凝血活酶時間(APTT)、瑞斯託黴素誘導的血小闆凝集試驗(RIPA)、血管性血友病因子(vWF)瑞斯託黴素輔因子(vWF∶RCo)、vWF抗原(vWF∶Ag)、vWF活性(vWF∶A)測定,vWF膠原結閤試驗(vWF∶CB),vWF與FⅧ結閤試驗(vWF∶FⅧ∶B)和多聚體分析對3例vWD先證者和傢繫成員進行錶型診斷.提取3例先證者外週血基因組DNA,用PCR法擴增vWF基因的52箇外顯子及側翼序列,通過直接測序分析vWF基因變異.結果 3例左證者APTT均延長,BT除先證者3外均正常,血漿RIPA、vWF∶RCo、vWF∶Ag、vWF∶A和vWF∶CB均有不同程度的減低.多聚體分析結果顯示先證者3中、高分子質量多聚體缺失,而先證者l、2基本正常.對3例先證者進行基因測序,共髮現3箇雜閤突變,分彆為vWF基因的G144067A(R2287Q)、G110374A(R1374H)、C110770T(S1506L)以及28號外顯子G110667A(D1472H)雜閤多態性.結論 R2287Q、R1374H和S1506L這3種雜閤錯義突變及D1472H雜閤多態性是分彆導緻3例先證者髮生遺傳性vWD的分子髮病機製.其中R2287Q是國際首次報道的新突變.
목적 대3개유전성혈관성혈우병(vWD)가계진행표형진단화기인형분석,탐토기분자발병궤제.방법 분별채용출혈시간(BT)、활화부분응혈활매시간(APTT)、서사탁매소유도적혈소판응집시험(RIPA)、혈관성혈우병인자(vWF)서사탁매소보인자(vWF∶RCo)、vWF항원(vWF∶Ag)、vWF활성(vWF∶A)측정,vWF효원결합시험(vWF∶CB),vWF여FⅧ결합시험(vWF∶FⅧ∶B)화다취체분석대3례vWD선증자화가계성원진행표형진단.제취3례선증자외주혈기인조DNA,용PCR법확증vWF기인적52개외현자급측익서렬,통과직접측서분석vWF기인변이.결과 3례좌증자APTT균연장,BT제선증자3외균정상,혈장RIPA、vWF∶RCo、vWF∶Ag、vWF∶A화vWF∶CB균유불동정도적감저.다취체분석결과현시선증자3중、고분자질량다취체결실,이선증자l、2기본정상.대3례선증자진행기인측서,공발현3개잡합돌변,분별위vWF기인적G144067A(R2287Q)、G110374A(R1374H)、C110770T(S1506L)이급28호외현자G110667A(D1472H)잡합다태성.결론 R2287Q、R1374H화S1506L저3충잡합착의돌변급D1472H잡합다태성시분별도치3례선증자발생유전성vWD적분자발병궤제.기중R2287Q시국제수차보도적신돌변.
Objective To analyze phenotype and genotype of three Chinese pedigrees with von Willebrand disease (vWD), and explore the molecular mechanism. Methods Bleeding time (BT),activated partial thromboplastin time ( APTT), ristocetin-induced platelet aggregation ( RIPA ), von Willebrand factor timer analysis were used for phenotype diagnosis. Genomic DNA was extracted from the peripheral blood (PB). All the 52 exons and flanking sequences of the probands' vWF gene were amplified by PCR and analyzed by direct sequencing. Results APTT were prolonged in all three probands, while BT were normal exferent extents. In multimer analysis, proband 3 lost the large and intermediate molecular weight multimers,while proband 1 and 2 were normal. Gene analysis in the three probands revealed three heterozygous missense mutations of 144067 G→A(R2287Q) in exon 39 , 110374G→A (R1374H)and 110770C→T(S1506L) in exon 28 and heterozygous polymorphism 110667G→A (D1472H) in exon 28, respectively. Conclusion The three heterozygous mutations (R2287Q,R1374H and S1506L) and an heterozygous polymorphism (D1472H) are genetic defects of the hereditary vWD of the three pedigrees respectively. R2287Q is a novel mutation reported for the first time in the literature.
