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Compatibility of stabilized whole blood products with CD4 technologies and their suitability for quality assessment programs.

Ding T, Bergeron M, Seely P, Yang X, Diallo TO, Plews M, Sandstrom P, Ball TB, Meyers AF - PLoS ONE (2014)

Bottom Line: This study demonstrated that the characteristics of commercially available SWBPs vary across multiple CD4 platforms.The compatibility of testing panels for EQA programs with multiple analytical platforms needs to be carefully considered, especially in large multiplatform CD4 EQA programs.The selection of a suitable cross-platform SWBP is an increasing challenge as more reagents and platforms are introduced for CD4 T-cell enumeration.

View Article: PubMed Central - PubMed

Affiliation: National Laboratory for HIV Immunology, Public Health Agency of Canada, Ottawa, Ontario, Canada.

ABSTRACT

Background: CD4 T cell enumeration is the most widely used prognostic marker for management of HIV disease. Internal quality control and external quality assessment (EQA) programs are critical to ensure reliability of clinical measurements. The utility of stabilized whole blood products (SWBP) as a test reagent for EQA programs such as Quality Assessment and Standardization for Immunological measures relevant to HIV/AIDS (QASI) program have been demonstrated previously. Since then, several new commercial SWBPs and alternative CD4 enumeration technologies have become available. Seven SWBPs were evaluated on seven different enumeration platforms to determine which product(s) are most suitable for EQA programs that support multiple analytical technologies.

Method: Assessment of SWBPs was based on two criteria: (1) accuracy of CD4 T cell measurements and; (2) stability under sub optimal storage conditions.

Results: Three SWBPs (Multi-Check, StatusFlow and CD4 Count) showed accurate CD4 T-cell absolute count and percentage values across six of the enumeration platforms. All products retain stability up to 18 days at 21-23°C with the exception of Multi-Check-high on FacsCount and Multi-Check-Low and StatusFlow-Low on Pima. One of the products (CD4 Count) retained stability for three days on all platforms tested when stored at 37°C.

Conclusion: This study demonstrated that the characteristics of commercially available SWBPs vary across multiple CD4 platforms. The compatibility of testing panels for EQA programs with multiple analytical platforms needs to be carefully considered, especially in large multiplatform CD4 EQA programs. The selection of a suitable cross-platform SWBP is an increasing challenge as more reagents and platforms are introduced for CD4 T-cell enumeration.

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Related in: MedlinePlus

Analysis of Low CD4 level SWBPs on CyFlow.Stabilized whole blood products (low CD4 level) and fresh whole blood stained with CD4% easy count on CyFlow Counter are shown. Each CD4×SSC dot plot displays two gates: (1) “CD4” gate set around CD4 lymphocytes cluster and (2) “LYM” gate set around all lymphocytes.
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pone-0103391-g002: Analysis of Low CD4 level SWBPs on CyFlow.Stabilized whole blood products (low CD4 level) and fresh whole blood stained with CD4% easy count on CyFlow Counter are shown. Each CD4×SSC dot plot displays two gates: (1) “CD4” gate set around CD4 lymphocytes cluster and (2) “LYM” gate set around all lymphocytes.

Mentions: We next assessed accuracy using CD4 T cell percentages, and found that again, the majority of SWBPs passed accuracy using CD4 T cell percentages (Table 6). However, accuracy failed with Immuno-Trol (High and Low), CD-Chex Plus BC-High, and CytoFix (High and Low) with residual values >3.0 using the FACSCount CD4 reagent kit and CD-Chex Plus (High and Low) was not measurable. Immuno-Trol-High and CytoFix (High and Low) failed on the Guava PCA using the Guava Auto CD4/CD4% kit. Immuno-Trol (High and Low) failed on the CyFlow Counter using the CD4% easy count kit. Figure 2 illustrates the analysis of SWBPs (low level) on the CyFlow Counter using the CD4% easy count reagent. The CD4×SSC dot plots displayed the CD4 and the lymphocyte gate. The resolution between CD4− lymphocyte and monocytes is critical for reliable gating for measurements of lymphocyte percentages. The resolution observed with Immuno-Trol was poor which increased the level of difficulty to draw a reliable gate and obtain high lymphocyte recovery and low monocyte contaminants. Thus, Multi-Check, StatusFlow and CD4 Count showed best accuracy and performance for both high and low CD4 percentage preparations on six of the platforms tested.


Compatibility of stabilized whole blood products with CD4 technologies and their suitability for quality assessment programs.

Ding T, Bergeron M, Seely P, Yang X, Diallo TO, Plews M, Sandstrom P, Ball TB, Meyers AF - PLoS ONE (2014)

Analysis of Low CD4 level SWBPs on CyFlow.Stabilized whole blood products (low CD4 level) and fresh whole blood stained with CD4% easy count on CyFlow Counter are shown. Each CD4×SSC dot plot displays two gates: (1) “CD4” gate set around CD4 lymphocytes cluster and (2) “LYM” gate set around all lymphocytes.
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4126665&req=5

pone-0103391-g002: Analysis of Low CD4 level SWBPs on CyFlow.Stabilized whole blood products (low CD4 level) and fresh whole blood stained with CD4% easy count on CyFlow Counter are shown. Each CD4×SSC dot plot displays two gates: (1) “CD4” gate set around CD4 lymphocytes cluster and (2) “LYM” gate set around all lymphocytes.
Mentions: We next assessed accuracy using CD4 T cell percentages, and found that again, the majority of SWBPs passed accuracy using CD4 T cell percentages (Table 6). However, accuracy failed with Immuno-Trol (High and Low), CD-Chex Plus BC-High, and CytoFix (High and Low) with residual values >3.0 using the FACSCount CD4 reagent kit and CD-Chex Plus (High and Low) was not measurable. Immuno-Trol-High and CytoFix (High and Low) failed on the Guava PCA using the Guava Auto CD4/CD4% kit. Immuno-Trol (High and Low) failed on the CyFlow Counter using the CD4% easy count kit. Figure 2 illustrates the analysis of SWBPs (low level) on the CyFlow Counter using the CD4% easy count reagent. The CD4×SSC dot plots displayed the CD4 and the lymphocyte gate. The resolution between CD4− lymphocyte and monocytes is critical for reliable gating for measurements of lymphocyte percentages. The resolution observed with Immuno-Trol was poor which increased the level of difficulty to draw a reliable gate and obtain high lymphocyte recovery and low monocyte contaminants. Thus, Multi-Check, StatusFlow and CD4 Count showed best accuracy and performance for both high and low CD4 percentage preparations on six of the platforms tested.

Bottom Line: This study demonstrated that the characteristics of commercially available SWBPs vary across multiple CD4 platforms.The compatibility of testing panels for EQA programs with multiple analytical platforms needs to be carefully considered, especially in large multiplatform CD4 EQA programs.The selection of a suitable cross-platform SWBP is an increasing challenge as more reagents and platforms are introduced for CD4 T-cell enumeration.

View Article: PubMed Central - PubMed

Affiliation: National Laboratory for HIV Immunology, Public Health Agency of Canada, Ottawa, Ontario, Canada.

ABSTRACT

Background: CD4 T cell enumeration is the most widely used prognostic marker for management of HIV disease. Internal quality control and external quality assessment (EQA) programs are critical to ensure reliability of clinical measurements. The utility of stabilized whole blood products (SWBP) as a test reagent for EQA programs such as Quality Assessment and Standardization for Immunological measures relevant to HIV/AIDS (QASI) program have been demonstrated previously. Since then, several new commercial SWBPs and alternative CD4 enumeration technologies have become available. Seven SWBPs were evaluated on seven different enumeration platforms to determine which product(s) are most suitable for EQA programs that support multiple analytical technologies.

Method: Assessment of SWBPs was based on two criteria: (1) accuracy of CD4 T cell measurements and; (2) stability under sub optimal storage conditions.

Results: Three SWBPs (Multi-Check, StatusFlow and CD4 Count) showed accurate CD4 T-cell absolute count and percentage values across six of the enumeration platforms. All products retain stability up to 18 days at 21-23°C with the exception of Multi-Check-high on FacsCount and Multi-Check-Low and StatusFlow-Low on Pima. One of the products (CD4 Count) retained stability for three days on all platforms tested when stored at 37°C.

Conclusion: This study demonstrated that the characteristics of commercially available SWBPs vary across multiple CD4 platforms. The compatibility of testing panels for EQA programs with multiple analytical platforms needs to be carefully considered, especially in large multiplatform CD4 EQA programs. The selection of a suitable cross-platform SWBP is an increasing challenge as more reagents and platforms are introduced for CD4 T-cell enumeration.

Show MeSH
Related in: MedlinePlus