评估单平台微量毛细管流式细胞仪用于检测HIV-1感染的泰国患者的绝对CD4+ T淋巴细胞计数

Evaluation of a single-platform microcapillary flow cytometer for enumeration of absolute CD4+ T-lymphocyte counts in HIV-1 infected Thai patients.

作者信息

Pattanapanyasat Kovit, Phuang-Ngern Yuwadee, Lerdwana Surada, Wasinrapee Punneeporn, Sakulploy Natthaga, Noulsri Egarit, Thepthai Charin, McNicholl Janet M

机构信息

Center of Excellence for Flow Cytometry, Office for Research and Development, Faculty of Medicine, Siriraj Hospital, Mahidol University, Bangkok, Thailand.

出版信息

Cytometry B Clin Cytom. 2007 Sep;72(5):387-96. doi: 10.1002/cyto.b.20167.

DOI:10.1002/cyto.b.20167

PMID:17474130

Abstract

BACKGROUND

Various assays are used to enumerate peripheral blood absolute CD4+ T-lymphocytes. Flow cytometry is considered the gold standard for this purpose. However, the high cost of available flow cytometers and monoclonal antibody reagents make it difficult to implement such methods in the resource-poor settings. In this study, we evaluated a cheaper, recently developed single-platform microcapillary cytometer for CD4+ T-lymphocyte enumeration, the personal cell analyzer (PCA), from Guava Technologies.

METHODS

CD4+ and CD8+ T-lymphocyte counts in whole blood samples from 250 HIV-1 infected Thais were determined, using a two-color reagent kit and the Guava PCA, and compared with the results obtained with two reference microbead-based methods from Becton Dickinson Biosciences: the three-color TruCOUNT tube method and the two-color FACSCount method. Statistical correlations and agreements were determined using linear correlation and Bland-Altman analysis.

RESULTS

Absolute CD4+ T-lymphocyte counts obtained using the Guava PCA method highly correlated with those obtained using TruCOUNT method (R(2) = 0.95, mean bias +13.1 cells/microl, limit of agreement [LOA]-117.9 to +144.1 cells/microl) and the FACSCount method (R2 = 0.94, mean bias = +33.2 cells/microl, LOA-101.8 to +168.3 cells/microl). Absolute CD8+ T-lymphocyte counts obtained using the Guava PCA method also highly correlated with those obtained with the two reference methods (R(2) = 0.92 and 0.88, respectively).

CONCLUSION

This study shows that the enumeration of CD4+ T-lymphocytes using the Guava microcapillary cytometer PCA method performed well when compared with the two reference bead-based methods. However, like the two reference methods, this new method needs substantial technical expertise.

摘要

背景

多种检测方法用于对外周血中绝对 CD4 + T 淋巴细胞进行计数。流式细胞术被认为是此目的的金标准。然而，现有流式细胞仪和单克隆抗体试剂成本高昂，使得在资源匮乏地区难以实施此类方法。在本研究中，我们评估了一种更便宜、最近开发的用于 CD4 + T 淋巴细胞计数的单平台微毛细管细胞仪——来自 Guava Technologies 的个人细胞分析仪（PCA）。

方法

使用双色试剂试剂盒和 Guava PCA 测定了 250 名感染 HIV - 1 的泰国人全血样本中的 CD4 + 和 CD8 + T 淋巴细胞计数，并与 Becton Dickinson Biosciences 两种基于微珠的参考方法所得结果进行比较：三色 TruCOUNT 管法和双色 FACSCount 法。使用线性相关和 Bland - Altman 分析确定统计相关性和一致性。

结果

使用 Guava PCA 方法获得的绝对 CD4 + T 淋巴细胞计数与使用 TruCOUNT 方法（R(2) = 0.95，平均偏差 +13.1 个细胞/微升，一致性界限[LOA] -117.9 至 +144.1 个细胞/微升）和 FACSCount 方法（R2 = 0.94，平均偏差 = +33.2 个细胞/微升，LOA -101.8 至 +168.3 个细胞/微升）获得的计数高度相关。使用 Guava PCA 方法获得的绝对 CD8 + T 淋巴细胞计数也与两种参考方法所得结果高度相关（分别为 R(2) = 0.92 和 0.88）。