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The comparability of the AQUIOS Tetra System was assessed by comparing the results with the predicate method using the estimation of difference as described in CLSI EP9-A3 Method Comparison and Bias Estimation Using Patient Samples. For CD3+, CD3+/CD4+, CD3+/CD8+, CD3-/(CD56+CD16+), and CD3-CD19+, the comparator method is samples stained with MultiTEST™ CD3-FITC/CD8-PE/CD45-PerCP/CD4-APC and MultiTEST™ CD3-FITC/CD16+CD56-PE/CD45-PerCP/CD19-APC monoclonal antibody reagents run on a BD FACSCalibur™ Flow Cytometer with MultiTEST™ 4-color TBNK software.

For CD45+ and CD45+ Low SS, the comparator method is specimens run on a UniCel DxH 800 hematology analyzer. Both method comparisons are comprised of 78% clinical specimens and 22% normal specimens. The data in Figure 1 through Figure 8 supports the premise that the systems are equivalent in their performance enumerating mature T, B, and NK lymphocytes in peripheral whole blood.

Values are expressed in terms of percentage (%) of the total lymphocyte count and as absolute count (cells/µL) for CD3+, CD3+/CD4+, CD3+/CD8+, CD3-/CD19+, and CD3-/CD56+ CD16+ cells as well as percentage (%) of CD45+ Low SS and absolute count (cells/µL) for CD45+ and CD45+Low SS when analyzed using Tetra Combo analytical mode.

Table 1. Method Comparison Between the AQUIOS Tetra Application and the BD FACSCalibur™

AQUIOS TETRA-1 PANEL CD45-FITC/CD4-RD1/CD8-ECD/CD3-PC5 (N=443)

Mean BD FACSCalibur™ Mean AQUIOS Tetra Correlation Slope Intercept

% + Lymphocytes

CD3+ 71.64 71.75 0.986 0.963 2.786

CD3+/CD4+ 31.11 31.26 0.996 0.997 0.228

CD3+/CD8+ 38.59 38.16 0.955 0.983 0.210

Cells/µL

CD3+ 1146 1118 0.977 0.978 10.453

CD3+/CD4+ 514 499 0.986 0.984 2.657

CD3+/CD8+ 603 582 0.982 0.965 4.689

AQUIOS TETRA-2+ PANEL CD45-FITC/CD56+CD16-RD1/CD19-ECD/CD3-PC5 (N=443)

% + Lymphocytes

CD3+ 71.65 71.93 0.987 0.970 2.433

CD3-/CD19+ 14.01 13.61 0.987 0.990 -0.260

CD3-/CD56+CD16+ 12.93 13.18 0.986 0.998 0.278

Cells/µL

CD3+ 1166 1106 0.973 0.950 9.643

CD3-/CD19+ 226 207 0.975 0.873 9.665

CD3-/CD56+CD16+ 190 181 0.979 0.966 2.756

COMPARING THE AQUIOS CL FLOW CYTOMETER, THE FACSCALIBUR™ FLOW CYTOMETER, AND THE UNICEL DXH 800 HEMATOLOGY ANALYZER TETRA APPLICATIONS

Visualize the Possibilities

Table 2. Method Comparison Between the AQUIOS Tetra Application and the UniCel DxH 800

AQUIOS TETRA-1 PANEL CD45-FITC/CD4-RD1/CD8-ECD/CD3-PC5 (N=445)

Mean UniCel DxH 800 Mean AQUIOS Tetra Correlation Slope Intercept

%

CD45+ Low SS 27.60 27.49 0.995 0.999 -0.084

Cells/µL

CD45+ 6131 5991 0.992 0.985 -51.430

CD45+ Low SS 1583 1537 0.993 0.977 -6.156

AQUIOS Tetra-1 Panel CD45-FITC/CD4-RD1/CD8-ECD/CD3-PC5

Figure 1. Regression Analysis for the CD3+ Cells using the AQUIOS Tetra Application and the BD FACSCalibur™

Figure 2. Regression Analysis for the CD3+/CD4+ Cells using the AQUIOS Tetra Application and the BD FACSCalibur™

Figure 3. Regression Analysis for the CD3+/CD8+ Cells using the AQUIOS Tetra Application and the BD FACSCalibur™

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Visualize the Possibilities

Figure 4. Regression Analysis for the CD45+ Cells using the AQUIOS Tetra Application and the UniCel DxH 800

Figure 5. Regression Analysis for the CD45+ Low SS Cells using the AQUIOS Tetra Application and the UniCel DxH 800

AQUIOS Tetra-2+ Panel CD45-FITC/(CD56+CD16)-RD1/CD19-ECD/CD3-PC5

Figure 6. Regression Analysis for the CD3+ Cells using the AQUIOS Tetra Application and the BD FACSCalibur™

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Figure 7. Regression Analysis for the CD3-/CD19+ Cells using the AQUIOS Tetra Application and the BD FACSCalibur™

Figure 8. Regression Analysis for the CD3-/CD56+CD16+ Cells using the AQUIOS Tetra Application and the BD FACSCalibur™

ConclusionsThe data demonstrates substantial equivalence of the AQUIOS CL Flow Cytometer to the FACSCalibur™ Flow Cytometer and the UniCel DxH 800 Hematology Analyzer.

Note: The cleared parameters of CD45 and CD45 low Side Scatter were not directly compared to the FACSCalibur™ Flow Cytometer since these parameters do not currently appear in the FACSCalibur™ Flow Cytometer intended use statement.

AQUIOS CL is a Class I Laser Product.

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FLOW-1131WP10.15-A

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