Abstract
| Original language | Undefined |
|---|---|
| Pages (from-to) | 397-407 |
| Number of pages | 11 |
| Journal | Cytometry. Part B : Clinical cytometry |
| Volume | 72B |
| Issue number | 5 |
| DOIs | |
| Publication status | Published - 2007 |
Keywords
- METIS-240634
- Single platform
- HIV monitoring
- Image cytometer
- CD4 enumeration
- IR-72257
- immunomagnetic separation
- immunofluorescent labeling
- point-of-care
Cite this
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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 journal › Article › Academic › peer-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 -