Qin Huan-huan, Wang Xue-feng, Ding Qiu-lan, Xu Guan-qun, Zhang Li-wei, Dai Jing, Lu Ye-ling, Xi Xiao-dong, Wang Hong-li
State Key Laboratory of Medical Genomics, Shanghai Institute of Hematology, Ruijin Hospital Affiliated to Shanghai Jiaotong University School of Medicine, Shanghai 200025, China.
Zhonghua Xue Ye Xue Za Zhi. 2011 Feb;32(2):99-102.
To analyze phenotype and genotype of three Chinese pedigrees with von Willebrand disease (vWD), and explore the molecular mechanism.
Bleeding time (BT), activated partial thromboplastin time (APTT), ristocetin-induced platelet aggregation (RIPA), von Willebrand factor (vWF): ristocetin cofactor (RCof) (vWF:RCof), vWF antigen (vWF:Ag), vWF activity (vWF:A) test, vWF collagen binding assay (vWF:CB), vWF and Factor VIII (FVIII) binding assay (vWF:FVIII:B) and multimer 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.
APTT were prolonged in all three probands, while BT were normal excepting for proband 3. Plasma RIPA, vWF:RCo, vWF:Ag, vWF:A and vWF:CB were decreased in different 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.
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.
分析3个中国血管性血友病（vWD）家系的表型和基因型，探讨其分子机制。
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聚集（RIPA）、血管性血友病因子（vWF）：瑞斯托霉素辅因子（RCof）（vWF:RCof）、vWF抗原（vWF:Ag）、vWF活性（vWF:A）检测、vWF胶原结合试验（vWF:CB）、vWF与凝血因子Ⅷ（FVIII）结合试验（vWF:FVIII:B）及多聚体分析进行表型诊断。从外周血（PB）中提取基因组DNA。先证者vWF基因的全部52个外显子及其侧翼序列经聚合酶链反应（PCR）扩增后直接测序分析。
3例先证者APTT均延长，除先证者3外BT均正常。血浆RIPA、vWF:RCo、vWF:Ag、vWF:A及vWF:CB均有不同程度降低。多聚体分析中，先证者3缺失大分子量和中等分子量多聚体，先证者1和2正常。3例先证者基因分析分别发现外显子39的144067G→A（R2287Q）、外显子28的110374G→A（R1374H）和110770C→T（S1506L）3个杂合错义突变以及外显子28的杂合多态性110667G→A（D1472H）。
3个家系遗传性vWD的遗传缺陷分别为3个杂合突变（R2287Q、R1374H和S1506L）及1个杂合多态性（D1472H）。R2287Q是文献首次报道的新突变。
