流式细胞术分析骨髓增生异常综合征小巨核细胞的初步研究

Study on micromegakaryocytes of patients with myelodysplastic syndrome by flow cytometry

目的建立基于巨核细胞糖蛋白Ⅱb/Ⅲa（CD41a）和碘化丙啶（PI）双标记的流式细胞术（FCM）分析骨髓小巨核细胞的方法,并研究其在骨髓增生异常综合征（MDS）诊断中的意义。方法采用异硫氰酸荧光素标记的单克隆抗体标记CD41a,PI标记巨核细胞DNA,用FCM分析巨核细胞倍体分布。结果 42例MDS患者,FCM检测小巨核细胞阳性率为90.5%,瑞氏-姬姆萨染色和免疫组织化学方法检测阳性率分别为54.8%和64.3%,三者比较差异有统计学意义（χ~2=13.640,P=0.001）;FCM检测MDS低危组和高危组小巨核细胞阳性率分别为81.8%和100%,差异有统计学意义（χ~2=4.019,P=0.045）。结论 FCM检测MDS小巨核细胞检出率高,且高危患者小巨核细胞检出率更高,有助于MDS的早期诊断和预后评估。

Objective To setup a measurement of human bone marrow micromegakaryocyte which based on CD41a and PI double-labeled flow cytometric analysis,and study the significance in the diagnosis of MDS. Methods In 42 cases of MDS patients, their bone marrow megakaryocytes were obtained by Percoll density gradient separation medium. The megakaryocyte glycoprotein Ⅱ b/Ⅲ a（CD41a）were marked with fluorescein isothiocyanate through its corresponding monoclonal antibody,and their DNA were marked with PI. Then the megakaryocyte ploidy was analyzed by flow cytometry（FCM）. Results The method for micromegakaryo- cyte identification and analysis was established. In 42 patients with MDS, the detection rate of micromegakaryocyte was 90.5 per- cent by FCM analysis, but only 54.8 percent by Wright-Giemsa staining test and 64.3 percent by immunohistochemistry, the differ- ence among them was statistically significant（χ2 = 13. 640, P=0. 001）. The 42 patients with MDS were divided into two groups （low-risk group and high-risk group）. The detection rates of mieromegakaryocyte were 81.8 percent in low-risk group and 100 per- cent in high-risk group separately by FCM analysis, the difference was statistically significant （χ2 =4.019, P = 0. 045）. Conclusion The detection rate of micromegakaryocyte by FCM with CD41a and PI double marker is higher than that by cytochemical staining. The detection rate of micromegakaryocyte in the high-risk group is higher than that of the low-risk group,which shows that the de- tection of micromegakaryocyte is of great significance for MDS prognosis assessment.