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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 different SWBPs.Multi-Check, StatusFlow and CD4 Count (low CD4 level) were prepared with MultiTest reagent CD3FITC/CD8PE/CD45PerCP/CD4APC on product incubated for 1 (D1) and 2 (D2) days at 37°C. Analysis displayed CD45×SSC dot plots with automated CD45 gates.
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pone-0103391-g003: FacsCalibur Multiset analysis of different SWBPs.Multi-Check, StatusFlow and CD4 Count (low CD4 level) were prepared with MultiTest reagent CD3FITC/CD8PE/CD45PerCP/CD4APC on product incubated for 1 (D1) and 2 (D2) days at 37°C. Analysis displayed CD45×SSC dot plots with automated CD45 gates.

Mentions: Incubation of SWBPs at suboptimal temperatures triggers sample degradation. Morphology and spectral properties may be lost rapidly. Testing of Multi-Check and StatusFlow products on the FacsCalibur was not continued beyond 2 days due to the inability to objectively gate the lymphocyte population as illustrated in Figure 3. The cursors placement around CD3 and CD4 cells clusters on Guava PCA was also challenging with StatusFlow and Multi-Check incubated a single day at 37°C, increasing the risk for unreliable measurements (Figure 4).


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 different SWBPs.Multi-Check, StatusFlow and CD4 Count (low CD4 level) were prepared with MultiTest reagent CD3FITC/CD8PE/CD45PerCP/CD4APC on product incubated for 1 (D1) and 2 (D2) days at 37°C. Analysis displayed CD45×SSC dot plots with automated CD45 gates.
© Copyright Policy
Related In: Results  -  Collection

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

pone-0103391-g003: FacsCalibur Multiset analysis of different SWBPs.Multi-Check, StatusFlow and CD4 Count (low CD4 level) were prepared with MultiTest reagent CD3FITC/CD8PE/CD45PerCP/CD4APC on product incubated for 1 (D1) and 2 (D2) days at 37°C. Analysis displayed CD45×SSC dot plots with automated CD45 gates.
Mentions: Incubation of SWBPs at suboptimal temperatures triggers sample degradation. Morphology and spectral properties may be lost rapidly. Testing of Multi-Check and StatusFlow products on the FacsCalibur was not continued beyond 2 days due to the inability to objectively gate the lymphocyte population as illustrated in Figure 3. The cursors placement around CD3 and CD4 cells clusters on Guava PCA was also challenging with StatusFlow and Multi-Check incubated a single day at 37°C, increasing the risk for unreliable measurements (Figure 4).

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