Integrated Biobank of Luxembourg, 1 rue Louis Rech, 3555 Dudelange, Luxembourg; International Society for Biological and Environmental Repositories (ISBER), Biospecimen Science Working Group, Canada.
Integrated Biobank of Luxembourg, 1 rue Louis Rech, 3555 Dudelange, Luxembourg.
J Immunol Methods. 2019 Feb;465:13-19. doi: 10.1016/j.jim.2018.11.012. Epub 2018 Nov 26.
Uncontrolled preanalytical variables can reduce the accuracy and reproducibility of downstream analytical results from peripheral blood mononuclear cells (PBMCs).
PBMCs were isolated from EDTA and citrate-anticoagulated blood samples, obtained from healthy subjects and patients with inflammatory and infectious conditions. PBMC-derived RNA samples were examined for gene expression changes induced by extended blood pre-centrifugation delays at 4 °C and RT. We used Taqman RTqPCR to evaluate the combination of two target genes for their "diagnostic performance" in identifying EDTA and citrate-anticoagulated PBMC samples with extended pre-centrifugation times.
We established the PBMC preanalytical score, a gene expression metric to asses the PBMC quality related to the pre-centrifugation delay at room temperature for different anticoagulants. The PBMC preanalytical score measurement can identify: CONCLUSION: The proposed PBMC preanalytical score may enable objective PBMC sample qualification for downstream applications, which may be influenced by blood precentrifugation delays.
未加控制的样本前处理变量会降低外周血单个核细胞（PBMC）下游分析结果的准确性和重现性。
从健康受试者和炎症性及感染性疾病患者的 EDTA 和柠檬酸盐抗凝血液样本中分离 PBMC。检测在 4°C 和 RT 下延长离心前处理时间对 PBMC 衍生 RNA 样本中基因表达变化的影响。我们使用 Taqman RTqPCR 评估两个靶基因的组合，用于鉴定 EDTA 和柠檬酸盐抗凝 PBMC 样本在延长离心前处理时间后的“诊断性能”。
我们建立了 PBMC 样本前处理评分，这是一种评估与不同抗凝剂室温下离心前处理时间相关的 PBMC 质量的基因表达指标。PBMC 样本前处理评分测量可以识别：
所提出的 PBMC 样本前处理评分可以对下游应用中可能受到血液离心前处理时间影响的 PBMC 样本进行客观的定性。
