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目的探讨流式细胞术(FCM)在诊断非霍奇金淋巴瘤(NHL)骨髓浸润中的价值。方法采用骨髓涂片和切片联合FCM检测NHL骨髓侵犯。骨髓涂片经瑞氏-吉姆萨液染色,分类计数瘤细胞。骨髓切片采用塑料包埋法制作组织切片,常规苏木精-姬姆萨-酸性品红(HGF)染色。流式细胞仪每份标本分析10 000个细胞。检测方案采用CD45/侧向散射角(SSC)设门,重点分析幼稚细胞群及成熟淋巴细胞群,经前向散射角(FSC)、SSC、异硫氰酸荧光素(FITC)标记的单克隆抗体(McAb1-FITC)、藻红蛋白(PE)标记的单克隆抗体(McAb2-PE)、CD45-Percp五参数分析确定免疫表型。结果 17例骨髓首诊病例中,骨髓穿刺(BMA)确诊12例(70.6%),骨髓活检(BMB)确诊15例(88.2%),FCM确诊17例(100%),FCM确诊率最高,较BMA差异有统计学意义(P<0.05),较BMB差异无统计学意义(P>0.05)。BMB检查发现NHL骨髓浸润方式间质型、结节型和弥漫型均易见,分别占23.5%,23.5%和52.9%。FCM检测结果为B细胞淋巴瘤13例(76.5%)、T细胞淋巴瘤和NK/T细胞淋巴瘤4例(23.5%)。结论骨髓涂片和骨髓切片检查是诊断NHL-BMI的基本方法,可以做出定性诊断。FCM可进一步做出明确的病理诊断。对于骨髓检查首诊NHL-BMI的病例,3种技术的联合应用可以提高NHL-BMI的诊断正确性和阳性率。
Objective To investigate the value of flow cytometry (FCM) in the diagnosis of bone marrow infiltration in non-Hodgkin’s lymphoma (NHL). Methods Bone marrow smears and sections combined with FCM were used to detect NHL bone marrow infiltration. Bone marrow smear by Wright - Giemsa staining, counting the number of tumor cells. Bone marrow sections were sectioned by plastic embedding and stained with hematoxylin-eosin (HGM). Flow cytometry analysis of 10 000 cells per sample. The detection protocol was based on CD45 / SSC, focusing on analysis of naive and mature lymphocyte populations, single clones labeled with FSC, SSC, and fluorescein isothiocyanate (FITC) Immunophenotypes were confirmed by five-parameter analysis of McAb1-FITC, PE-McAb (McAb2-PE) and CD45-Percp. Results In the first case of bone marrow biopsy, bone marrow biopsy (BMA) was confirmed in 12 cases (70.6%), bone marrow biopsy (BMB) was confirmed in 15 cases (88.2%) and FCM was diagnosed in 17 cases (100%). BMA had statistical significance (P <0.05), but no significant difference compared with BMB (P> 0.05). BMB examination found interstitial, nodular and diffuse type of NHL bone marrow infiltration were 23.5%, 23.5% and 52.9% respectively. FCM showed 13 cases (76.5%) of B-cell lymphoma, 4 cases (23.5%) of T-cell lymphoma and NK / T cell lymphoma. Conclusion Bone marrow smears and bone marrow biopsy are the basic methods to diagnose NHL-BMI, which can make qualitative diagnosis. FCM can further make a clear pathological diagnosis. For the first diagnosis of NHL-BMI bone marrow examination cases, the combination of three techniques can improve the diagnostic accuracy and positive rate of NHL-BMI.