Evaluation of a quantitative determination of CD4 and CD8 molecules as an alternative to CD4 <b>+</b> and CD8 <b>+</b> T lymphocyte counts in Africans

Diagbouga, Serge; Durand, Georges; Sanou, P T; Dahourou, H; Ledru, Éric

doi:10.1046/j.1365-3156.1999.00362.x

Cited by 22 publications

(15 citation statements)

References 16 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…This problem has been discussed in various publications [e.g. [8][9][10][11][12], and proposals have been made for progresses toward CD4 T-cell counting protocols which can be handled in remote areas at low costs [12][13][14].…”

Section: Discussionmentioning

confidence: 99%

A New No-Lyse, No-Wash Flow-Cytometric Method for the Determination of CD4 T Cells in Blood Samples

Greve¹,

Cassens²,

Westerberg³

et al. 2003

Transfus Med Hemother

View full text Add to dashboard Cite

Background: Commonly used flow-cytometric methods for immunophenotyping are based on erythrocyte lysing reagents. It is known from the literature that these reagents result in a significant loss of leucocytes caused by membrane destruction. Although the dual-platform method should compensate this phenomenon, the subset-specific individual differences in sensitivity to lysing reagents lead to incorrect values. In order to overcome this problem we introduce a no-lyse, no-wash procedure in combination with absolute true volumetric counting (TVC) for the enumeration of CD4 T cells. Material and Methods: Whole-blood samples of 50 blood donors and 20 samples of patients with acquired immunodeficiency syndrome (AIDS) were treated with both a lyse, no-wash and a nolyse, no-wash procedure. Then, CD4 T cells were counted with a TVC flow cytometer (CyFlow Counter). 30 blood samples of blood donors were treated with a lyse and wash protocol and measured by the CyFlow Counter and FACS-Calibur system (reference method). A new gating strategy was used for the data analysis for both, the lyse and no-lyse method and compared to the traditional gating procedure. Results: The TVC method showed a good reproducibility for both, the lyse (CV 1.9%) and the no-lyse procedure (CV 1.5%). CD4 counts measured by the no-lyse procedure are on average 10% higher than by using the lysing protocol. The comparison of the CyFlow Counter and FACS-Calibur results showed a correlation of r = 0.961. The simplified gating strategy shows a good correlation to the traditional gating procedure (r = 0.998). Conclusion: Erythrocyte lysing procedures cause substantial cell loss with individual values for every single subclass and patient. Therefore, the use of a no-lyse procedure is recommended. The new gating strategy is fully comparable to the traditional one and simplifies enumeration of CD4 T cells. Using the no-lyse procedure in combination with the new gating strategy, it is possible to reduce the costs per sample from EUR 30.– to below EUR 2.–.

show abstract

Section: Discussionmentioning

confidence: 99%

A New No-Lyse, No-Wash Flow-Cytometric Method for the Determination of CD4 T Cells in Blood Samples

Greve¹,

Cassens²,

Westerberg³

et al. 2003

Transfus Med Hemother

View full text Add to dashboard Cite

show abstract

“…21,22 In the four studies examining enzyme imd d munoassay, correlations between CD4 cell count as measured by dualdplatform flow cytometry and enzyme immunod d assay were all r <0.70. 12,21,23,24 When a blood fixative was employed using dualdplatform flow cytometry with pand d leucogating, correlations were r = 0.97 at day 0, r = 0.98 between days 0 and 3, and r = 0.92 between days 0 and 7.…”

Section: Clinical Informationmentioning

confidence: 99%

Diagnostic tests in HIV management: a review of clinical and laboratory strategies to monitor HIV-infected individuals in developing countries

Kimmel

Losina

Freedberg

et al. 2006

Bull World Health Organ

View full text Add to dashboard Cite

We conducted a systematic review on the performance of diagnostic tests for clinical and laboratory monitoring of HIVinfected adults in developing countries. Diagnostic test information collected from computerized databases, bibliographies and the Internet were categorized as clinical (non-laboratory patient information), immunologic (information from immunologic laboratory tests), or virologic (information from virologic laboratory tests). Of the 51 studies selected for the review 28 assessed immunologic tests, 12 virologic tests and seven clinical and immunologic tests. Methods of performance evaluation were primarily sensitivity and specificity for the clinical category and correlation coefficients for immunologic and virologic categories. In the clinical category, the majority of test performance measures was reported as >70% sensitive and >65% specific. In the immunologic category, correlation coefficients ranged from r = 0.54 to r = 0.99 for different CD4 count enumeration techniques, while correlation for CD4 and total lymphocyte counts was between r = 0.23 and r = 0.74. In the virologic category, correlation coefficients for different human immunodeficiency virus (HIV) ribonucleic acid (RNA) quantification techniques ranged from r = 0.54 to r = 0.90. Future research requires consensus on designing studies, and collecting and reporting data useful for decision-makers. We recommend classifying information into clinically relevant categories, using a consistent definition of disease across studies and providing measures of both association and accuracy.

show abstract

“…While FACS machines are commonly accessible in developed countries, its high equipment and operation costs have limited their uses in resource-limited areas where most of the HIV-infected subjects reside. On the other hand, magnetic-activated cell sorting (MACS) technique has also been utilized for CD4 T cell counting 6,7 as an attempt to bring down the in initial equipment cost to make CD4 T cell testing more affordable. In contrast to FACS, which interrogates samples on a particle-by-particle basis, MACS involves the mixing of the sample with magnetic beads that have been attached with antibodies or other molecules that recognize the surface marker on the target cell.…”

Section: Introductionmentioning

confidence: 99%

Fast sorting of CD4+ T cells from whole blood using glass microbubbles

et al. 2015

View full text Add to dashboard Cite

The isolation of CD4 positive T lymphocyte (CD4+) from peripheral blood is important for monitoring patients after HIV infection. Here, we demonstrate a fast isolation strategy for CD4+ cells that involves mixing blood and glass microbubbles. After the specific binding of target cells to the microbubbles carrying specific antibodies on their surface, target cells will spontaneously float to the top of the blood vial and can be quickly separated. Using this strategy, we demonstrate that the isolation of CD4+ cells in less than 5 minutes and with better than 90% efficiency. This strategy for cell isolation based on buoyancy and glass microbubbles is quick and inexpensive, minimizes blood handling, does not require magnetic fields, or centrifugation equipment, and could lead to new, efficient strategies for AIDS diagnosis in resource-limited areas.

show abstract

Evaluation of a quantitative determination of CD4 and CD8 molecules as an alternative to CD4 + and CD8 + T lymphocyte counts in Africans

Cited by 22 publications

References 16 publications

A New No-Lyse, No-Wash Flow-Cytometric Method for the Determination of CD4 T Cells in Blood Samples

A New No-Lyse, No-Wash Flow-Cytometric Method for the Determination of CD4 T Cells in Blood Samples

Diagnostic tests in HIV management: a review of clinical and laboratory strategies to monitor HIV-infected individuals in developing countries

Fast sorting of CD4+ T cells from whole blood using glass microbubbles

Contact Info

Product

Resources

About