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目的寻找骨髓造血功能障碍性疾病中巨细胞病毒(CMV)活动性感染的组织学依据。方法华中科技大学同济医学院附属协和医院2006年2月至2007年10月筛选造血系统两系或三系减少患儿37例(再生障碍性贫血26例,骨髓增生异常综合征11例)。取患儿骨髓活检组织,塑料包埋制备薄切片;将切片置于Eppen-dorf管反应液内进行原位逆转录并扩增,原位检测骨髓组织中CMV mRNA表达。同时取患儿骨髓液行普通液相RT-PCR检测CMV mRNA作为对照。结果14例(37.8%)骨髓塑料切片检测到CMV mRNA表达;阳性细胞多为骨髓实质细胞,呈局灶性或片状分布,阳性信号主要位于细胞胞浆内,少量出现在部分细胞的胞核。普通液相RT-PCR检测16例(43.2%)骨髓液中有CMV mRNA表达,2种方法阳性率差异无统计学意义(P>0.05)。结论改良的原位RT-PCR方法可用于塑料包埋切片的检测,可定位诊断骨髓中CMV活动性感染,为CMV活动性感染致血液学改变提供了组织学依据,对骨髓造血功能障碍性疾病发病机制的探讨具有重要意义。
OBJECTIVE: To investigate the histological basis of cytomegalovirus (CMV) active infection in bone marrow hematopoietic disorders. Methods A total of 37 children with aplastic anemia (aplastic anemia in 26 cases and 11 cases with myelodysplastic syndromes) were screened from February 2006 to October 2007 in Huazhong University of Science and Technology Affiliated Union Hospital of Tongji Medical College. Bone marrow biopsy specimens were taken from children and embedded in plastic for thin sections. The sections were placed in Eppen-dorf tube and reversely transcribed and amplified in situ. CMV mRNA expression in bone marrow was detected in situ. At the same time take the bone marrow of children with normal liquid phase RT-PCR detection of CMV mRNA as a control. Results CMV mRNA expression was detected in bone marrow of 14 cases (37.8%). Most of the positive cells were bone marrow parenchyma cells with focal or patchy distribution. The positive signals mainly located in the cytoplasm of the cells and a small amount appeared in the nuclei of some cells . The expression of CMV mRNA in bone marrow of 16 cases (43.2%) was detected by ordinary liquid-phase RT-PCR. There was no significant difference between the two methods (P> 0.05). Conclusion The modified in situ RT-PCR method can be used for the detection of plastic-embedded sections. It can be used to diagnose active CMV infection in bone marrow and provide a histological basis for hematological changes caused by active CMV infection. The improved method of hematopoietic disorders The pathogenesis of the discussion is of great significance.