Severe von Willebrand disease due to a defect at the level of von Willebrand factor mRNA expression: detection by exonic PCR-restriction fragment length polymorphism analysis.
Severe von Willebrand disease due to a defect at the level of von Willebrand factor mRNA expression: detection by exonic PCR-restriction fragment length polymorphism analysis.
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DOI：
10.1073/pnas.88.9.3857
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发表时间：
1991-05
影响因子：
11.1
通讯作者：
William C. Nichols;Susan E. Lyons;Jonathon S. Harrison;Robert L. Cody;David Ginsburg
中科院分区：
文献类型：
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作者：
William C. Nichols;Susan E. Lyons;Jonathon S. Harrison;Robert L. Cody;David Ginsburg
血管性血友病（vWD）是人类最常见的遗传性出血性疾病，由血浆凝血蛋白血管性血友病因子（vWF）的异常引起。重度（III型）vWD是常染色体隐性遗传，与极低或检测不到的vWF水平相关。我们报告了一种方法，旨在区分mRNA表达的两个vWF等位基因的外周血血小板RNA的PCR分析，使用位于vWF基因的外显子内的DNA序列多态性。这种方法被应用于一个严重的vWD家系，其中三个八兄弟姐妹受到影响，父母和其他兄弟姐妹是临床正常的。每一个亲本都携带一个在mRNA水平上沉默的vWF等位基因。遗传两个异常等位基因的家庭成员受到严重vWD的影响，而只有一个异常等位基因的个体是无症状的。母系和父系沉默等位基因在两个编码序列多态性以及内含子40可变数目串联重复序列上是相同的，这表明可能有共同的起源。鉴于本文报告的两种外显子多态性的频率，该分析应适用于约70%的I型和III型vWD患者。这种比较DNA和RNA PCR-限制性片段长度多态性的方法也可能被证明是有用的，在确定与其他遗传疾病相关的基因表达水平的缺陷。
von Willebrand disease (vWD), the most common inherited bleeding disorder in humans, results from abnormalities in the plasma clotting protein von Willebrand factor (vWF). Severe (type III) vWD is autosomal recessive in inheritance and is associated with extremely low or undetectable vWF levels. We report a method designed to distinguish mRNA expression from the two vWF alleles by PCR analysis of peripheral blood platelet RNA using DNA sequence polymorphisms located within exons of the vWF gene. This approach was applied to a severe-vWD pedigree in which three of eight siblings are affected and the parents and additional siblings are clinically normal. Each parent was shown to carry a vWF allele that is silent at the mRNA level. Family members inheriting both abnormal alleles are affected with severe vWD, whereas individuals with only one abnormal allele are asymptomatic. The maternal and paternal silent alleles are identical at two coding sequence polymorphisms as well as an intron 40 variable number tandem repeat, suggesting a possible common origin. Given the frequencies of the two exon polymorphisms reported here, this analysis should be applicable to approximately 70% of type I and type III vWD patients. This comparative DNA and RNA PCR-restriction fragment length polymorphism approach may also prove useful in identifying defects at the level of gene expression associated with other genetic disorders.