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Use of CD45 gating in three and four-color flow cytometric immunophenotyping: guideline from the National Institute of Allergy and Infectious Diseases, Division of AIDS.在三色和四色流式细胞术免疫表型分析中使用CD45设门:美国国立过敏与传染病研究所艾滋病司指南
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Multisite comparison of CD4 and CD8 T-lymphocyte counting by single- versus multiple-platform methodologies: evaluation of Beckman Coulter flow-count fluorospheres and the tetraONE system. The NIAID DAIDS New Technologies Evaluation Group.单平台与多平台方法对CD4和CD8 T淋巴细胞计数的多中心比较:贝克曼库尔特流式细胞计数荧光微球和tetraONE系统的评估。美国国立过敏与传染病研究所艾滋病司新技术评估组
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采用双平台方法评估T细胞亚群的时间和温度稳定性。

Time and temperature stability of T-cell subsets evaluated by a dual-platform method.

作者信息

Olteanu Horatiu, Schur Bernard C, Harrington Alexandra M, Kroft Steven H

出版信息

Am J Blood Res. 2012;2(2):128-35. Epub 2012 May 25.

PMID:22762032
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC3384401/
Abstract

INTRODUCTION

T-cell subset enumeration in HIV patients is routinely performed for monitoring infection stage and response to antiretroviral therapy. Studies have examined the effect of specimen refrigeration and age for single-platform (SP) methods, but there is limited data for time and temperature requirements of dual-platform (DP) methods.

METHODS

Using a DP method, we analyzed peripheral blood (PB) from 52 HIV patients at room temperature (RT) at 24, 72, and 96 hours. PBs from 34 HIV patients had baseline RT analysis within 24 hours, and then were refrigerated and analyzed at 24, 48, and 72 hours. The coefficient of variation (CV) and residuals (changes in lymphocyte subsets) were recorded at each time point and compared to assess the precision and bias under the various conditions. Testing performance under different conditions was compared by linear regression.

RESULTS

Mean CV was ≤7.3% and median residuals were <30/μl for absolute CD4 and CD8 determinations. There was good correlation between baseline analysis data at RT and at various time points, both at RT and 4°C.

CONCLUSIONS

Our results are similar to those published for SP methods for aging or refrigerated specimens. The high level of agreement between measurements supports the robustness of this DP methodology.

摘要

引言

对HIV患者的T细胞亚群进行计数是监测感染阶段和抗逆转录病毒治疗反应的常规操作。已有研究探讨了标本冷藏和年龄对单平台(SP)方法的影响,但关于双平台(DP)方法的时间和温度要求的数据有限。

方法

我们采用双平台方法,在室温(RT)下于24、72和96小时对52例HIV患者的外周血(PB)进行分析。34例HIV患者的PB在24小时内进行基线RT分析,然后冷藏,并在24、48和72小时进行分析。记录每个时间点的变异系数(CV)和残差(淋巴细胞亚群变化),并进行比较,以评估不同条件下的精密度和偏差。通过线性回归比较不同条件下的检测性能。

结果

对于绝对CD4和CD8测定,平均CV≤7.3%,残差中位数<30/μl。RT下的基线分析数据与RT和4°C下不同时间点的数据之间存在良好的相关性。

结论

我们的结果与已发表的关于老化或冷藏标本的SP方法的结果相似。测量之间的高度一致性支持了这种双平台方法的稳健性。