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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

FacsCalibur Multiset analysis of stabilized whole blood products (SWBPs).Low CD4 level SWBP and fresh whole blood stained with CD4FITC/CD8PE/CD3PerCP antibody combination are shown. Two dot plots are shown for each analysis: CD3×CD4 with attractor gate on CD3+4− cells cluster; CD4×CD8 (upper right corner) with attractor gate on beads, CD4, CD8 and double positive CD4+8+ cells cluster.
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pone-0103391-g001: FacsCalibur Multiset analysis of stabilized whole blood products (SWBPs).Low CD4 level SWBP and fresh whole blood stained with CD4FITC/CD8PE/CD3PerCP antibody combination are shown. Two dot plots are shown for each analysis: CD3×CD4 with attractor gate on CD3+4− cells cluster; CD4×CD8 (upper right corner) with attractor gate on beads, CD4, CD8 and double positive CD4+8+ cells cluster.

Mentions: For absolute count measurements, we found with the following exceptions that the majority of the seven SWBPs passed the accuracy test (Table 5). CD-Chex Plus (High and Low) was not measurable on the FacsCount when the FacsCount reagent kit was used; CytoFix-Low was not measurable on the FacsCount when the FacsCount CD4 reagent kit was used. Accuracy failed with CD-Chex Plus-Low, CD-Chex plus BC-Low and CytoFix-Low with differences greater than 15% as compared to the reference values on the FacsCalibur using MultiSet software with the CD4/CD8/CD3 combination. Figure 1 illustrates the MultiSet analysis of SWBPs (low CD4 level) on the FacsCalibur using the CD4/CD8/CD3 TriTest reagent. Compared to fresh whole blood, resolution between CD3+4− and CD3+4+ cells populations was lower for all products. Poor resolution was also observed with CD-Chex Plus, CD-Chex Plus BC and CytoFix. CD-Chex Plus BC (High and Low) and CytoFix-High failed on the Guava PCA platform using the Guava Express CD3/CD4 reagent kit. CD-Chex Plus-Low failed on Guava PCA using the CD4/CD4% reagent. CD-Chex Plus BC-Low and Immuno-Trol-Low failed on CyFlow Counter when the CD4% easy count kit was used. Thus, Multi-Check, StatusFlow, and CD4 Count show best scoring performance for both high and low CD4 level preparations.


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)

FacsCalibur Multiset analysis of stabilized whole blood products (SWBPs).Low CD4 level SWBP and fresh whole blood stained with CD4FITC/CD8PE/CD3PerCP antibody combination are shown. Two dot plots are shown for each analysis: CD3×CD4 with attractor gate on CD3+4− cells cluster; CD4×CD8 (upper right corner) with attractor gate on beads, CD4, CD8 and double positive CD4+8+ cells cluster.
© Copyright Policy
Related In: Results  -  Collection

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

pone-0103391-g001: FacsCalibur Multiset analysis of stabilized whole blood products (SWBPs).Low CD4 level SWBP and fresh whole blood stained with CD4FITC/CD8PE/CD3PerCP antibody combination are shown. Two dot plots are shown for each analysis: CD3×CD4 with attractor gate on CD3+4− cells cluster; CD4×CD8 (upper right corner) with attractor gate on beads, CD4, CD8 and double positive CD4+8+ cells cluster.
Mentions: For absolute count measurements, we found with the following exceptions that the majority of the seven SWBPs passed the accuracy test (Table 5). CD-Chex Plus (High and Low) was not measurable on the FacsCount when the FacsCount reagent kit was used; CytoFix-Low was not measurable on the FacsCount when the FacsCount CD4 reagent kit was used. Accuracy failed with CD-Chex Plus-Low, CD-Chex plus BC-Low and CytoFix-Low with differences greater than 15% as compared to the reference values on the FacsCalibur using MultiSet software with the CD4/CD8/CD3 combination. Figure 1 illustrates the MultiSet analysis of SWBPs (low CD4 level) on the FacsCalibur using the CD4/CD8/CD3 TriTest reagent. Compared to fresh whole blood, resolution between CD3+4− and CD3+4+ cells populations was lower for all products. Poor resolution was also observed with CD-Chex Plus, CD-Chex Plus BC and CytoFix. CD-Chex Plus BC (High and Low) and CytoFix-High failed on the Guava PCA platform using the Guava Express CD3/CD4 reagent kit. CD-Chex Plus-Low failed on Guava PCA using the CD4/CD4% reagent. CD-Chex Plus BC-Low and Immuno-Trol-Low failed on CyFlow Counter when the CD4% easy count kit was used. Thus, Multi-Check, StatusFlow, and CD4 Count show best scoring performance for both high and low CD4 level preparations.

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