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

Research output: Contribution to journalArticleAcademicpeer-review

15 Citations (Scopus)

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.
Original languageUndefined
Pages (from-to)397-407
Number of pages11
JournalCytometry. Part B : Clinical cytometry
Volume72B
Issue number5
DOIs
Publication statusPublished - 2007

Keywords

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

Cite this

@article{3153fd47afe34d98ad3c6b9c493fbc38,
title = "CD4+T Lymphocytes enumeration by an easy-to-use single platform image cytometer for HIV monitoring in resource-constrained settings",
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.",
keywords = "METIS-240634, Single platform, HIV monitoring, Image cytometer, CD4 enumeration, IR-72257, immunomagnetic separation, immunofluorescent labeling, point-of-care",
author = "X. Li and Aurel Ymeti and Bj{\"o}rn Lunter and Christian Breukers and Tibbe, {Arjan G.J.} and Terstappen, {Leonardus Wendelinus Mathias Marie} and Jan Greve",
year = "2007",
doi = "10.1002/cyto.b.20165",
language = "Undefined",
volume = "72B",
pages = "397--407",
journal = "Cytometry. Part B : Clinical cytometry",
issn = "1552-4949",
publisher = "Wiley-Liss Inc.",
number = "5",

}

CD4+T Lymphocytes enumeration by an easy-to-use single platform image cytometer for HIV monitoring in resource-constrained settings. / Li, X.; Ymeti, Aurel; Lunter, Björn; Breukers, Christian; Tibbe, Arjan G.J.; Terstappen, Leonardus Wendelinus Mathias Marie; Greve, Jan.

In: Cytometry. Part B : Clinical cytometry, Vol. 72B, No. 5, 2007, p. 397-407.

Research output: Contribution to journalArticleAcademicpeer-review

TY - JOUR

T1 - CD4+T Lymphocytes enumeration by an easy-to-use single platform image cytometer for HIV monitoring in resource-constrained settings

AU - Li, X.

AU - Ymeti, Aurel

AU - Lunter, Björn

AU - Breukers, Christian

AU - Tibbe, Arjan G.J.

AU - Terstappen, Leonardus Wendelinus Mathias Marie

AU - Greve, Jan

PY - 2007

Y1 - 2007

N2 - 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.

AB - 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.

KW - METIS-240634

KW - Single platform

KW - HIV monitoring

KW - Image cytometer

KW - CD4 enumeration

KW - IR-72257

KW - immunomagnetic separation

KW - immunofluorescent labeling

KW - point-of-care

U2 - 10.1002/cyto.b.20165

DO - 10.1002/cyto.b.20165

M3 - Article

VL - 72B

SP - 397

EP - 407

JO - Cytometry. Part B : Clinical cytometry

JF - Cytometry. Part B : Clinical cytometry

SN - 1552-4949

IS - 5

ER -