CAJ | 학술논문

目的探索中国正常女性X-连锁葡萄糖6-磷酸脱氢酶(G6PD)、P55、BTK、FHL-1基因外显子多态性位点的杂合子基因型频率,并探讨应用以上位点采用基于X染色体失活克隆性检测法-逆转录检测法进行造血细胞克隆性分析的价值.方法采集446名中国正常女性外周血标本,提取基因组DNA,应用等位基因特异性PCR或PCR-限制性片段长度多态性方法分析X-连锁G6PD、P55、BTK、FHL-1基因外显子多态性位点的杂合基因型频率.以上述杂合位点作为克隆性分析的标记,采用逆转录检测法分析有JAK2V617F突变或克隆性染色体异常的原发性血小板增多症(ET)女性患者造血细胞的克隆性.结果 446名中国正常女性G6PD基因杂合基因型频率为12.8%(57例),P55基因杂合基因型频率为29.4%(131例),BTK基因杂合基因型频率为52.0%(232例),FHL-1基因杂合基因型频率为46.4%(207例).4个多态性位点中具有至少1个杂合位点的正常女性占81.4%(363例).对G6PD、P55、BTK或FHL-1为杂合子且伴JAK2V617F突变的10例ET患者,应用逆转录检测法进行造血细胞克隆性分析,所有患者均为单克隆/寡克隆.结论基于X染色体失活的克隆性检测法-逆转录检测法可用于约80%中国女性患者的克隆性分析.
목적 탐색중국정상녀성X-련쇄포도당6-린산탈경매(G6PD)、P55、BTK、FHL-1기인외현자다태성위점적잡합자기인형빈솔,병탐토응용이상위점채용기우X염색체실활극륭성검측법-역전록검측법진행조혈세포극륭성분석적개치.방법 채집446명중국정상녀성외주혈표본,제취기인조DNA,응용등위기인특이성PCR혹PCR-한제성편단장도다태성방법 분석X-련쇄G6PD、P55、BTK、FHL-1기인외현자다태성위점적잡합기인형빈솔.이상술잡합위점작위극륭성분석적표기,채용역전록검측법분석유JAK2V617F돌변혹극륭성염색체이상적원발성혈소판증다증(ET)녀성환자조혈세포적극륭성.결과 446명중국정상녀성G6PD기인잡합기인형빈솔위12.8%(57례),P55기인잡합기인형빈솔위29.4%(131례),BTK기인잡합기인형빈솔위52.0%(232례),FHL-1기인잡합기인형빈솔위46.4%(207례).4개다태성위점중구유지소1개잡합위점적정상녀성점81.4%(363례).대G6PD、P55、BTK혹FHL-1위잡합자차반JAK2V617F돌변적10례ET환자,응용역전록검측법진행조혈세포극륭성분석,소유환자균위단극륭/과극륭.결론 기우X염색체실활적극륭성검측법-역전록검측법가용우약80%중국녀성환자적극륭성분석.
Objective 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). Methods 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. Results 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. Conclusion Clonality detection based on X chromosome inactivation patterns - transcription based clonality assays is applicable to about 80% of Chinese females.