CD4+ T lymphocytes enumeration by an easy-to-use single platform image cytometer for HIV monitoring in resource-constrained settings

Li, Xiao and Ymeti, Aurel and Lunter, Björn and Breukers, Christian and Tibbe, Arjan G.J. and Terstappen, Leon W.M.M. and Greve, Jan (2007) CD4+ T lymphocytes enumeration by an easy-to-use single platform image cytometer for HIV monitoring in resource-constrained settings. Cytometry Part B: Clinical Cytometry, 72 (5). pp. 397-407. ISSN 1552-4949

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Abstract:	Backround: HIV monitoring in resource-constrained settings demands affordable and reliable CD4+ T lymphocytes enumeration methods. We developed a simple single platform image cytometer (SP ICM), which is a dedicated volumetric CD4+ T lymphocytes enumeration system that uses immunomagnetic and immunofluorescent technologies. The instrument was designed to be a low-cost, yet reliable and robust one. In this article we test the instrument and the immunochemical procedures used on blood from HIV negative and HIV positive patients. - Methods: After CD4 immunomagnetic labeling in whole blood, CD4+ T lymphocytes, CD4+dim monocytes and some nonspecifically labeled cells are magnetically attracted to an analysis surface. Combining with CD3-Phycoerythrin (PE) labeling, only CD3+CD4+ T lymphocytes are fluorescently labeled and visible in a fluorescent image of the analysis surface. The number of CD4+ T lymphocytes is obtained by image analysis. Alternatively, CD3 immunomagnetic selection in combination with CD4 immunofluorescent labeling can also be applied for CD4+ T lymphocytes enumeration. - Results: The SP ICM system was compared with two single platform flow cytometer (SP FCM) methods: tetraCXP and TruCount methods. The SP ICM system has excellent precision, accuracy and linearity for CD4+ T lymphocytes enumeration. Good correlations were obtained between the SP ICM and the SP FCM methods for blood specimens of 44 HIV- patients, and of 63 HIV+ patients. Bland-Altman plots showed interchangeability between the SP ICM and the SP FCM methods. - Conclusions: The immunolabeling methods and the instrumentation are simple and easy-to-handle for less-trained operators. The SP ICM system is a good candidate for CD4+ T lymphocytes enumeration in point-of-care settings of resource-constrained countries.
Item Type:	Article
Copyright:	© 2007 Wiley InterScience
Faculty:	Science and Technology (TNW)
Research Group:	Medical Cell BioPhysics (MCBP)
Link to this item:	http://purl.utwente.nl/publications/72257
Official URL:	http://dx.doi.org/10.1002/cyto.b.20165
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