摘要
目的 对一个遗传性凝血因子V缺乏症家系进行凝血因子V(FV)基因突变的检测。方法 经用活化部分凝血活酶时间(APTT),凝血酶原时间(PT)及FV促凝活性(FV:C)和FV抗原(FV:Ag)测定进行表型诊断;用PCR法对先证者(女,16岁)的FV基因25个外显子及其侧翼序列进行扩增,PCR产物纯化后直接测序,检测其基因突变。突变位点经限制性内切酶分析证实。108名健康献血者作对照。结果 先证者APTT 126.6 s,PT42.8 s,FⅤ:C0.3％,FⅤ:Ag1.3％,FⅡ:C、FⅤⅡ:C、FⅤⅢ:C、FⅨ:C、FⅩ:C和Fbg均在正常范围内;FⅤ外显子区共发现5个与GeneBank Z99572序列不同的位点,其中突变位点为位于第8外显子区的G1348T和位于第14外显子区的4887～8delG。家系分析表明前者遗传于父亲,后者遗传于母亲。结论 G1348T引起的错义突变和4887～8delG引起的移码突变,是导致先证者FⅤ缺乏的原因。这是2个导致遗传性FⅤ缺乏症的新的FⅤ基因突变位点。
Objective To discover the gene mutations of a pedigree with inherited factor V ( FV) deficiency. Methods The activated partial thromboplastin time (APTT) , prothrombin time (PT) , FV activity (FV:C) and FV antigen test were adopted for phenotype diagnosis. The genomic DNA was extracted from the peripheral blood of the 16-year-old propositus, female. All the 25 exons and their flanks in the FV gene of the propositus were amplified by polymerase chain reaction (PCR) . The PCR products were screened by direct sequencing and the mutations were further confirmed by restricted enzyme digestion. Six persons in the pedigree (grandfather, grandmother, father, mother, uncle, and aunt) were examined too. 108 healthy blood donors were used as controls. Results The APTT ,PT ,FV:C, and FV:Ag of the propositus were 126.6s,42.8s,0.3% and 1.3% respectively. The Fbg and F Ⅱ , FV Ⅱ , FV Ⅲ , FIX, FX activities were in normal range. FV: C of the members of the pedigree was 36% -70% , and the FV: Ag of the pedigree members was 26.4% - 45.3% that of the mixture of 30 normal plasma samples. Taking the GeneBank Z99572 sequence as the reference, totally five variations in the FV gene were found in the propositus. The mutations, A1348G and 4887 - SdelG, were traced to her father and her mother respectively. No 1348 G→T mutation was found in the 108 controls. Conclusion The FV deficiency of the propositus is caused by missense mutation of G1348T and frameshift mutation of 4887 -8delG, which haven't been identified previously.
出处
《中华医学杂志》
CAS
CSCD
北大核心
2003年第4期312-315,共4页
National Medical Journal of China
