检验医学 ›› 2015, Vol. 30 ›› Issue (1): 21-25.DOI: 10.3969/j.issn.1673-8640.2015.01.006

• 临床应用研究·论著 • 上一篇    下一篇

慢性淋巴细胞白血病CD5漏检临床案例分析及对策

唐古生, 柳敏, 胡晓霞, 高磊, 杨建民, 王健民   

  1. 上海市长海医院血液内科,上海 200433
  • 收稿日期:2014-02-26 出版日期:2015-01-30 发布日期:2015-02-02
  • 作者简介:null

    作者简介:唐古生,男,1978年生,博士,主治医师,主要研究方向为血液系统恶性疾病免疫学临床诊断。

  • 基金资助:
    国家自然科学基金面上项目(81270638);国家自然科学基金青年基金资助项目(81102249)

Analysis on the missing detection of CD5 in chronic lymphocytic leukemia and its countermeasures

TANG Gusheng, LIU Min, HU Xiaoxia, GAO Lei, YANG Jianmin, WANG Jianmin.   

  1. Department of Hematology, Changhai Hospital, Shanghai 200433, China
  • Received:2014-02-26 Online:2015-01-30 Published:2015-02-02

摘要:

目的 分析不同荧光素标记抗体对慢性淋巴细胞白血病(CLL)B细胞表面CD5检测结果差异及其对临床诊断的影响。方法 采用流式细胞仪检测3例CD5表达强度不同的慢性淋巴细胞白血病(CLL)患者的免疫表型,采用SYSMEX XE2100 全自动血液分析仪检测血常规;以1例急性B淋巴细胞白血病(B-ALL)患者作为阴性对照。结果 采用异硫氰酸荧光素(FITC)标记抗体检测4例患者骨髓异常B淋巴细胞CD5表达,CD5阳性细胞未见独立成群,阳性细胞比例分别为47.1%、17.7%、6.7%和7.9%;以≥20%为标准,仅病例1阳性。以PE-Cy7荧光素标记抗体重新检测,病例1和2的CD5阳性细胞独立成群,病例3 CD5表达呈连续表达模式,阳性细胞比例分别上升至91.1%、70.3%和38.2%。而对照病例4(B-ALL患者)2次检测CD5比例均为阴性。病例1、2、3符合典型CLL免疫表型。结论 CLL患者B细胞表面常异常表达CD5,但其表达强度显著低于T细胞表面CD5表达。采用弱荧光素标记抗体检测B细胞表面CD5表达,会出现漏检进而影响临床诊断。对弱表达抗原选择合适的荧光素标记,并进行验证和比对确定其准确性,是正确提供疾病免疫表型的重要保障。

关键词: CD5, 免疫表型, 慢性淋巴细胞性白血病, 流式细胞仪, 漏检

Abstract:

Objective To analyze the variation of CD5 caused by different fluorescein-labeled antibodies on B cells in patients with chronic lymphocytic leukemia(CLL) and the influence on clinical diagnosis. Methods Flow cytometry and SYSMEX XE2100 automated hematology analyzer were used for the detection of immunophenotype and blood cell counts of 3 CLL patients with different expression intensities of CD5, respectively. 1 patient with B-cell acute lymphocytic leukemia(B-ALL) was selected as negative control. Result In 4 patients with abnormal B lymphocytes, using fluorescein isothiocyanate(FITC) fluorescein-labeled antibody, no independent groups expressing CD5 were identified, and the proportions of CD5 positive cells were 47.1%, 17.7%, 6.7% and 7.9%. For ≥20% as standard, there was only 1 positive case. Re-tested with PE-Cy7 fluorescein-labeled antibody, however, there were independent groups of CD5 positive cells in case 1 and 2, and there was still a continuous expression pattern of CD5 in case 3. The proportions of CD5 positive cells increased to 91.1%, 70.3% and 38.2% in these 3 specimens from CLL patients. In case 4, the specimen from B-ALL patient, the proportions of CD5 positive cells were negative in the 2 conditions. Case 1, 2 and 3 were typical CLL phenotyping. Conclusions CD5 is often abnormally expressed on B cells in CLL patients, but its expression intensity is significantly lower than that on T cells. Using weak fluorescein-labeled antibodies might miss CD5 expression on B cells, and thus it will affect the clinical diagnosis because of leak detection. It should select appropriate fluorescein-labeled antibodies for weakly expressed antigens and conduct validation and comparison to determine its accuracy, which is an important safeguard for the provision of correct disease phenotype.

Key words: CD5, Immunophenotype, Chronic lymphocytic leukemia, Flow cytometry, Leak detection

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