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

X. Li, Aurel Ymeti, Björn Lunter, Christian Breukers, Arjan G.J. Tibbe, Leonardus Wendelinus Mathias Marie Terstappen, Jan Greve

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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.
Original languageUndefined
Pages (from-to)397-407
Number of pages11
JournalCytometry. Part B : Clinical cytometry
Issue number5
Publication statusPublished - 2007


  • METIS-240634
  • Single platform
  • HIV monitoring
  • Image cytometer
  • CD4 enumeration
  • IR-72257
  • immunomagnetic separation
  • immunofluorescent labeling
  • point-of-care

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