应用DNA测序技术检测血友病B患者FⅨ基因突变  

Detection of Factor Ⅸ gene Mutation in Paitients with Hemophilia B by DNA Sequencing

作　　者：张媛[1] 杨林花[1] 陆晔玲[2] 丁秋兰[2] 王学锋[2] 刘秀娥[1] 侯丽虹[1] 

机构地区：[1]山西医科大学第二医院血液科,山西太原030001 [2]上海第二医科大学附属瑞金医院输血科,上海200025

出　　处：《中国实验血液学杂志》2009年第2期476-478,共3页Journal of Experimental Hematology

基　　金：山西省科技厅攻关项目(编号022073);山西省自然基金项目(编号04107)

摘　　要：为了研究3名中国汉族非亲缘系血友病B患者凝血因子Ⅸ(FⅨ)基因突变的类型,常规检测患者血浆APTT及FⅨ∶C,进行表型诊断;抽提患者外周血白细胞基因组DNA,应用PCR技术分8段扩增FⅨ基因外显子序列,用双脱氧终止法检测核酸序列。结果表明:与正常对照者相比,HB患者APTT测定值明显升高,FⅨ∶C测定值明显降低,PT测定值正常。测序结果显示3例HB患者均有相应的基因序列改变,患者1有外显子6G22119A点突变,患者2有外显子1G7932C点突变,患者3有外显子T32685C点突变。结论:3例HB患者均检测到相应的基因序列改变,这为HB患者基因缺陷的分子机制提供了证据。In order to investigate the patterns of FⅨ gene mutation in 3 unrelated hemophilia B （HB） patients, the activated partial thromboplastin time （APTT） and FⅨ activity （FⅨ： C） tests were adopted for phenotype diagnosis. All of the eight exons and their flank of FⅨ gene were amplified by polymerase chain reaction（ PCR）, the nucleic acid sequences were detected by dideoxymediated chain-termination method. The results indicated that as compared with normal control, the APTT value significantly increased, F Ⅸ：C value obviously decreased, PT value was normal. Sequencing results showed that all of 3 HB patients had the changes of gene sequences, among 3 patients the G22119A point mutation of exon 6 existed in case No. 1, the G7932C point mutation of exon 2 was detected in case No. 2 and the T32685C point mutation of exon 8 was found in case No. 3. In conclusion, the relevant changes of gene sequences in all of 3 HB patients were detected, which provides some evidences for molecular mechanism of gene deficiency in HB patients.

关 键 词：血友病B 凝血因子Ⅸ 基因突变 DNA测序技术 

分 类 号：R554.1[医药卫生—血液循环系统疾病]