UKNEQAS for leucocyte immunophenotyping: the first 10 years
UKNEQAS for leucocyte immunophenotyping: the first 10 years
复制标题
DOI：
10.1136/jcp.54.7.508
复制
发表时间：
2001-07-01
影响因子：
3.4
通讯作者：
Barnett, D
中科院分区：
文献类型：
--
作者：
Reilly, JT;Barnett, D
在过去的十年里，细胞免疫表型已经成为诊断血液学和免疫学中的一门新学科，在白血病的快速诊断和监测人类免疫缺陷病毒感染者的疾病进展方面具有非常重要的价值。台式流式细胞仪的引入意味着免疫表型现在也用于CD34(+)外周血干细胞(PBSCs)的定量，以确保造血祖细胞采集的正确时机和充分性。此外，流式细胞仪已成为白细胞去除后血液成分中白细胞计数的重要工具。由于这一新学科现在是临床领域的主要诊断和预后工具，它的使用必须受到内部和外部质量控制的约束。早在1986年，这种要求就被首次认识到，当时为使用碱性磷酸酶抗碱性磷酸酶技术进行白血病免疫细胞化学诊断的实验室启动了区域间质量评估计划(IRQAS)。该项目始于大约25个英国实验室。1990年，在引入了另外两个项目(一个是使用紫外线显微镜和后来的流式细胞仪诊断白血病，另一个是CD4(+)T细胞的计数)后，IRQAS获得了英国国家外部质量评估计划(UK NEQAS)的资格，并将其名称改为UK NEQAS for Leuccell免疫表型。在过去的十年中，曾经很小的IRQAS计划已经发展成为同类计划中最大的国际计划，为世界各地650多个实验室提供了EQA，用于白血病免疫表型、淋巴细胞亚群分析、外周血干细胞，以及最近的低水平白细胞计数。多年来，这一EQA计划突出了一些重要问题，如荧光染料和抗体滴度的不当使用，以及有效的门控策略的确定，所有这些都直接导致了细胞免疫表型鉴定中出现的实验室间高度差异。此外，特别是在淋巴细胞亚群、外周血干细胞的绝对计数和低数量白细胞的计数方面，英国白细胞免疫表型方案NEQAS有助于突出单平台和双平台流式细胞术技术之间的差异。由于这些发现，英国用于白细胞免疫表型鉴定的NEQAS有助于减少实验室间的差异，并使英国和国际上的标准化程度更高。这些进展归功于英国NEQAS用于白细胞免疫表型鉴定的独特的全血稳定过程，该过程确保保留细胞所需的物理特征(光散射和抗原性)，以确保成功的细胞免疫表型鉴定。这一重大技术进步使用于EQA目的的标本能够在全球范围内分发，具有最小的基质影响，并在稳定后的几个月内表现为与新鲜血液相同的方式。
In the past decade, cellular immunophenotyping has become a new discipline in diagnostic haematology and immunology, and is invaluable in the rapid diagnosis of leukaemia and monitoring disease progression in human immunodeficiency virus infected individuals. The introduction of bench top flow cytometers has meant that immunophenotyping is now also used for the quantitation of CD34(+) peripheral blood stem cells (PBSCs) to ensure the correct timing and adequacy of haematopoietic progenitor cell harvests. Furthermore, flow cytometry has become an important tool for the counting of leucocytes in blood components after leucocyte depletion. Because this new discipline is now such a major diagnostic and prognostic tool in the clinical arena, its use must be subject to both internal and external quality control. Such a requirement was first recognised as early as 1986 when an Inter-Regional Quality Assessment Scheme (IRQAS) was initiated for laboratories that undertook the immunocytochemical diagnosis of leukaemia using the alkaline phosphates antialkaline phosphatase technique. This programme began with around 25 UK laboratories. In 1990, after the introduction of two more programmes (one for leukaemia diagnosis using UV microscopy and latterly flow cytometry, and one for the enumeration of CD4(+) T cells) the IRQAS achieved UK National External Quality Assessment Scheme (UK NEQAS) status and changed its title to UK NEQAS for Leucocyte Immunophenotyping. In the past decade the once small IRQAS programme has evolved into the largest international scheme of its kind, providing EQA to over 650 laboratories world wide for leukaemia immunophenotyping, lymphocyte subset analysis, PBSCs, and more recently low level leucocyte counting. Over the years, this EQA programme has highlighted important problems, such as the inappropriate use of fluorochromes and antibody titre, and the identification of effective gating strategies, all of which have contributed directly to the high interlaboratory variations seen in cellular immunophenotyping. Furthermore, particularly in absolute counting of lymphocyte subsets, PBSCs, and the enumeration of low numbers of leucocytes, UK NEQAS for Leucocyte Immunophenotyping programmes have been instrumental in highlighting the differences that occur between single and dual platform flow cytometric technologies. As a result of these findings, UK NEQAS for Leucocyte Immunophenotyping has helped to reduce the variation seen on an interlaboratory basis and enabled greater standardisation both in the UK and internationally. These advances have been attributable to the development, by UK NEQAS for Leucocyte Immunophenotyping, of a unique whole blood stabilising process that ensures the retention of the physical characteristics (both light scatter and antigenic profile) required of cells to ensure successful cellular immunophenotyping. This major technological advancement has enabled the distribution of specimens for EQA purposes on a global scale that have minimal matrix effect and behave in a manner identical to fresh blood for several months after stabilisation.