Liu Liu, Ma Xiao-tang, Wang Jie-yu, Su Tao, Yang Lin, Xu Ze-feng, Qin Tie-jun, Xiao Zhi-jian
Institute of Hematology and Blood Diseases Hospital, CAMS & PUMC, Tianjin, China.
Zhonghua Xue Ye Xue Za Zhi. 2012 Apr;33(4):261-5.
To explore the frequencies of heterozygosity in X-linked G6PD, P55, BTK, and FHL-1 gene exonic polymorphic loci among Chinese females and the value of determination of hematopoietic clonality by detection of these X-chromosome exonic polymorphisms based on X-chromosome inactivation patterns (XCIP)-transcription-based clonality assays (TCA).
Genomic DNA was extracted from peripheral blood of 446 Chinese healthy females. Allele-specific PCR (ASPCR) or PCR-restriction enzyme digestion method was applied for detecting G6PD, P55, BTK and FHL-1 polymorphisms. Those heterozygotic loci were used as markers to examine the hematopoietic clonality of bone marrow mononuclear cells by TCA from essential thrombocythemia (ET) patients with JAK2V617F mutation and myelodysplastic syndrome (MDS) patients with abnormal karyotype.
Among the total 446 genomic DNA samples, the frequencies of heterozygosity in G6PD, P55, BTK and FHL-1 loci were 12.8%, 29.4%, 52.0% and 46.4%, respectively. About 81.4% of females were heterozygous at one or more loci. All 10 ET patients with JAK2V617F mutation and 2 MDS patients with abnormal karyotype, which were heterozygotic in either locus, had monoclonal/oligoclonal hematopoiesis.
Clonality detection based on X chromosome inactivation patterns-transcription based clonality assays is applicable to about 80% of Chinese females.
探讨中国女性中X连锁葡萄糖-6-磷酸脱氢酶(G6PD)、P55、布鲁顿酪氨酸激酶(BTK)和FHL-1基因外显子多态性位点的杂合频率,以及基于X染色体失活模式(XCIP)-转录克隆性分析(TCA)检测这些X染色体外显子多态性来确定造血克隆性的价值。
从446名中国健康女性外周血中提取基因组DNA。采用等位基因特异性PCR(ASPCR)或PCR-限制性内切酶消化法检测G6PD、P55、BTK和FHL-1基因多态性。以这些杂合位点为标记,通过TCA检测携带JAK2V617F突变的原发性血小板增多症(ET)患者和核型异常的骨髓增生异常综合征(MDS)患者骨髓单个核细胞的造血克隆性。
在446份基因组DNA样本中,G6PD、P55、BTK和FHL-1位点的杂合频率分别为12.8%、29.4%、52.0%和46.4%。约81.4%的女性在一个或多个位点呈杂合状态。10例携带JAK2V617F突变的ET患者和2例核型异常的MDS患者,无论在哪个位点呈杂合状态,均存在单克隆/寡克隆造血。
基于X染色体失活模式-转录克隆性分析的克隆性检测适用于约80%的中国女性。