一种用于血清学检测和T细胞亚群免疫表型分析的联合HIV-1蛋白微珠阵列，与混合流式细胞仪联用：迈向用于传染病诊断和监测的综合多任务仪器平台的一步。

A combined HIV-1 protein bead array for serology assay and T-cell subset immunophenotyping with a hybrid flow cytometer: a step in the direction of a comprehensive multitasking instrument platform for infectious disease diagnosis and monitoring.

作者信息

Faucher Sylvie, Martel Alexandre, Sherring Alice, Bogdanovic Dragica, Malloch Laurie, Kim John E, Bergeron Michèle, Sandstrom Paul, Mandy Francis F

机构信息

National HIV Immunology Laboratory, National HIV and Retrovirology Laboratories, Public Health Agency of Canada, Ottawa, Ontario.

出版信息

Cytometry B Clin Cytom. 2006 May;70(3):179-88. doi: 10.1002/cyto.b.20109.

DOI:10.1002/cyto.b.20109

PMID:16615079

Abstract

BACKGROUND

A new generation of bench-top flow cytometers with digital signal processing to perform suspension array technology (SAT) based bead array assays as well as leukocyte immunophenotyping is now available. These hybrid instruments provide an opportunity for the development of a more cost effective multitasking platform to support infectious disease treatment in resource limited countries.

METHODS

We report the development and testing of two modules compatible with the hybrid flow cytometers. The first module is an eleven HIV-1 protein bead array (PBA) for the detection of circulating antibodies and the second is a cell based T-cell enumeration assay.

RESULTS

The HIV-1 PBA was tested in parallel with two enzyme immunoassays (EIAs) for the detection of plasma antibodies from 4 HIV-1 seroconversion panels and a low antibody titer panel. The PBA as well as the two EIAs performed equally for the detection of antibody positive samples from all seroconversion panels. One antibody positive sample from the low antibody titer panel was missed by the PBA together with one of the two EIAs tested. A parallel analysis of the HIV-1 PBA with Western blot (a confirmatory test for HIV infection) using plasma from nine HIV-1(+) individuals showed that the HIV-1 PBA detected more of the gp41 and gp120 antibody positive samples. Preliminary CD4 T-cell immunophenotyping results from 14 HIV(+) and 10 HIV(-) whole blood specimens with the hybrid flow cytometer platform compared well to conventional flow cytometry data.

CONCLUSION

The successful combination of bead and cell based assays on a single hybrid instrument demonstrated the potential utility of a multitasking platform. The results presented are providing groundwork for future development of more cost effective modular architecture for a flexible flow cytometry based platform.

摘要

背景

新一代具备数字信号处理功能的台式流式细胞仪现已问世，可用于进行基于悬浮阵列技术（SAT）的微珠阵列分析以及白细胞免疫表型分析。这些混合型仪器为开发更具成本效益的多任务平台提供了契机，以支持资源有限国家的传染病治疗。

方法

我们报告了与混合型流式细胞仪兼容的两个模块的开发与测试情况。第一个模块是用于检测循环抗体的11种HIV-1蛋白微珠阵列（PBA），第二个模块是基于细胞的T细胞计数分析。

结果

HIV-1 PBA与两种酶免疫测定法（EIA）并行测试，用于检测来自4个HIV-1血清转化样本组和一个低抗体滴度样本组的血浆抗体。在检测所有血清转化样本组中的抗体阳性样本时，PBA以及两种EIA的表现相当。PBA与所测试的两种EIA之一均遗漏了低抗体滴度样本组中的一个抗体阳性样本。使用9名HIV-1（+）个体的血浆对HIV-1 PBA与蛋白质印迹法（HIV感染的确证试验）进行的平行分析表明，HIV-1 PBA检测到更多的gp41和gp120抗体阳性样本。使用混合型流式细胞仪平台对14份HIV（+）和10份HIV（-）全血样本进行的初步CD4 T细胞免疫表型分析结果与传统流式细胞术数据相比良好。