CAJ | 학술논문

目的分析6例非肌性肌球蛋白重链9(MYH9)基因相关疾病患者临床特征和基因.方法取患者外周血,光镜下计数血小板数目、涂片并瑞氏染色后观察血小板形态以及有无中性粒细胞包涵体;抽提患者外周血基因组DNA,PCR扩增MYH9基因的40个外显子及两端侧翼序列,DNA测序并与基因库序列进行比对以确定基因异常,用限制性核酸内切酶和聚丙烯酰胺凝胶电泳法分析排除多态性.结果 6例患者血小板计数下降、体积增大,中性粒细胞内可见淡蓝色包涵体.基因分析发现在1号、30号和40号外显子中存在T97C(W33R)、4335InsCAGAAGAAG(1445InsQKK)、G4269A(D1424N)和G5833T(E1945Stop)4种基因突变,其中前2种突变为首次发现,并排除基因多态性的可能.结论 6例MYH9相关疾病患者的基因突变中T97C(W33R)和4335InsCAGAAGAAG(1445InsQKK)是国际上首次发现的新突变.病史较长且治疗反应不佳的原发免疫性血小板减少症患者应考虑MYH9基因相关疾病的可能性.
목적 분석6례비기성기구단백중련9(MYH9)기인상관질병환자림상특정화기인.방법 취환자외주혈,광경하계수혈소판수목、도편병서씨염색후관찰혈소판형태이급유무중성립세포포함체;추제환자외주혈기인조DNA,PCR확증MYH9기인적40개외현자급량단측익서렬,DNA측서병여기인고서렬진행비대이학정기인이상,용한제성핵산내절매화취병희선알응효전영법분석배제다태성.결과 6례환자혈소판계수하강、체적증대,중성립세포내가견담람색포함체.기인분석발현재1호、30호화40호외현자중존재T97C(W33R)、4335InsCAGAAGAAG(1445InsQKK)、G4269A(D1424N)화G5833T(E1945Stop)4충기인돌변,기중전2충돌변위수차발현,병배제기인다태성적가능.결론 6례MYH9상관질병환자적기인돌변중T97C(W33R)화4335InsCAGAAGAAG(1445InsQKK)시국제상수차발현적신돌변.병사교장차치료반응불가적원발면역성혈소판감소증환자응고필MYH9기인상관질병적가능성.
Objective To investigate clinical features and to identify gene mutations in six patients with nonmuscle myosin heavy chain 9 gene (MYH9)-related disease. Methods The platelet counts were measured using automated complete blood cell counter and manual manner. The size of platelets and inclusion bodies were observed under light microscopy. All the 40 exons and exon-intron boundaries of MYH9 gene were amplified by PCR and then DNA sequencing was performed. Restriction endonuclease analysis and polyacrylamide gel electrophoresis (PAGE)were used for polymorphism analysis. Results Six patients all shared the commom feasures of thrombocytopenia with giant platelets and granulocyte inclusions. Four MYH9 gene mutations were found in the six patients: T97C(W33R)in exon 1, 4335Insert CAGAAGAAG(1445InsQKK) and G4269A(D1424N)in exon 30 and G5833T(E1945Stop)in exon 40. The former two were novel mutations which have not been reported in the literature. The results of restriction endonuclease analysis and PAGE could exclude the possibility of nucleotide polymorphisms. Conclusions The MYH9 gene mutations were identified in six patients with MYH9 related disorders,and T97C(W33R)and 4335InsCAGAAGAAG(1445InsQKK)were novel mutations. MYH9 related disease should be considered in individuals with persistent thrombocytopenia which is non-responsive to corticosteroids and immuno-repressive agents.