Department of Immunopathology, SA Pathology at Women's and Children's Hospital, North Adelaide, SA, Australia; Adelaide Medical School, School of Biomedicine and the Robinson Research Institute, Faculty of Health Science, University of Adelaide, Adelaide, SA, Australia.
Department of Immunopathology, SA Pathology at Women's and Children's Hospital, North Adelaide, SA, Australia; Adelaide Medical School, School of Biomedicine and the Robinson Research Institute, Faculty of Health Science, University of Adelaide, Adelaide, SA, Australia; School of Biological Sciences, Faculty of Science, University of Adelaide, Adelaide, SA, Australia.
Pathology. 2024 Jun;56(4):571-576. doi: 10.1016/j.pathol.2023.11.011. Epub 2024 Feb 7.
Medical diagnostic laboratories have come under further scrutiny to ensure quality standards of their service and external quality assurance (EQA) programs involving multiple laboratories have been used to gauge this quality based on a consensus. However, because of the geographical distances within a country or internationally, cell surface marker expressions may change due to time delays and transport temperatures. Attention was given to this issue some decades ago and hence requires a re-evaluation in consideration of updated methods, reagents and instruments for flow cytometry and phenotyping. We have undertaken an extensive study to examine the effects of various conditions on blood storage akin to that experienced by patient samples as well as EQA programs, examining expression of lymphocyte surface markers, CD3, CD4, CD8, CD2, CD19, CD20, CD16/56 and HLA-DR. Assessment of lithium-heparin anticoagulated whole blood showed an increase in percentage of CD3 and CD8 T cells and a decrease in CD16/56 NK cells after storage at room temperature (RT) for 24 and/or 48 h. In comparison, storage at 4°C led to a decrease in percentage of CD4 and increase in percentage of CD8 cells. The low temperature also caused an increase in percentage of B cells (CD19, CD20). While storage at RT did not alter levels of HLA-DR CD3 T cells, there was a significant increase in percentage of these cells after 48 h. Changes were also seen at both temperatures when EDTA was used as an anti-coagulant. Assessment of blood treated with a stabiliser, normally used in the EQA samples (Streck Cell Preservative), reduced the range of lymphocyte subsets affected, with only CD2 and CD20 cells being significantly different at both temperatures, We conclude that 24-48 h storage/transport can affect the percentage of CD3, CD4 T cells, CD8 T cells, B cells, NK cells and HLADR T cells which can be minimised by using the blood stabiliser as per EQA programs and we emphasise the need to adopt this in the processing of patients' blood samples.
医学诊断实验室受到了进一步的审查，以确保其服务的质量标准，并且涉及多个实验室的外部质量保证（EQA）计划已经被用来根据共识来衡量这种质量。然而，由于国家内部或国际上的地理距离，细胞表面标志物的表达可能会由于时间延迟和运输温度而发生变化。几十年前就已经注意到了这个问题，因此需要重新评估，以考虑到更新的方法、试剂和仪器，用于流式细胞术和表型分析。我们进行了一项广泛的研究，以检查各种条件对血液储存的影响，类似于患者样本所经历的条件，以及 EQA 计划，检查淋巴细胞表面标志物、CD3、CD4、CD8、CD2、CD19、CD20、CD16/56 和 HLA-DR 的表达。评估锂肝素抗凝全血，结果表明，在室温（RT）下储存 24 小时和/或 48 小时后，CD3 和 CD8 T 细胞的百分比增加，而 NK 细胞（CD16/56）的百分比减少。相比之下，在 4°C 下储存导致 CD4 细胞的百分比降低，CD8 细胞的百分比增加。低温还导致 B 细胞（CD19、CD20）的百分比增加。虽然 RT 下储存不会改变 HLA-DR CD3 T 细胞的水平，但在 48 小时后，这些细胞的百分比显著增加。当 EDTA 用作抗凝剂时，在两种温度下也观察到了变化。评估使用稳定剂（通常用于 EQA 样本的稳定剂）处理的血液，减少了受影响的淋巴细胞亚群的范围，只有 CD2 和 CD20 细胞在两种温度下都有显著差异，我们得出结论，24-48 小时的储存/运输会影响 CD3、CD4 T 细胞、CD8 T 细胞、B 细胞、NK 细胞和 HLA-DR T 细胞的百分比，使用血液稳定剂可以将其最小化，这符合 EQA 计划，我们强调需要在处理患者的血液样本时采用这种方法。
