流式细胞术在首诊非霍奇金淋巴瘤骨髓侵犯中的作用

Effects of flow cytometry in bone marrow involvement of non-Hodgkin's lymphoma at initial diagnosis

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 research the value of flow cytometry（FCM） to examine bone marrow involvement（BMI） of non- Hodgkin＇s lymphoma（NHI.）. Methods Combined examination of BMA,BMB and FCM was used to detect the BMI of NHL. Bone marrow smears were stained by Wright Giemsa and tumor cells of NHL were differentially counted under optical microscope. BMB tissue was embedded in plastic. Sections were routinely stained with HGF. 10,000 cells were tested on each sample by FCM. Detec- tion scheme included designing CD45/SSC doors and analyzing importantly the group of immature-cell and mature lymphocyte. The five-parameter analysis including FSC, SSC, McAbl-FITC, McAh2-PE, CD45-Perep were used to determine the immunopheno- type. Results 12 cases of NHL-BMI were diagnosed by BMA（70.6%） ,15 cases by BMB（88.2%） and 17 cases by FCM（100%） in all 17 cases at initial diagnosis. The rate of diagnosis of FCM was higher than BMA and BMB,obviously different with BMA（P〈 0.05） and not different with BMB（P〉0.05）. Many involvement types of NHL-BMI were found in BMB,which the ratio of interstitial type was 23.5% ,nodular type was 23.5% and diffused type was 52.9% respectively. FCM result was 13 cases of B-cell lymphoma（76.5 % ）and 4 cases of T cell lymphoma and NK cell lymphoma. Conclusion At initial diagnosis of N HL,examination with BMA and BMB is a basic method and examination with FCM can provide precise views of the pathological classification further. Combined examination with BMA,BMB and FCM detecting BMI of NHL could improve the diagnostic accuracy and positive ratio.