Immunophenotypic Diagnosis of Lymphoproliferative Disorders Using the Abbott CELL-DYN Sapphire Hematology Analyzer.

The CELL-DYN Sapphire (Abbott) is a routine hematology analyzer which uses integrated optical and fluorescence measurements, with FL1 (FITC) and FL2 (PE) detectors being configured for fluorescence analysis. Enumeration of CD3/CD4 and CD8 lymphocytes as well as platelets quantification using an anti-CD61 monoclonal antibody (moAb) are now included in the automated process of the analyzer. Recently, Johannessen et al. (Clin.Lab.Haematol.,2006) demonstrated that the CELL-DYN Sapphire analyzer allows a possible complete automated leukocyte immunodifferential count, using a panel of different moAbs (Johannessen et al., 2006).

Aim To test the ability of this blood analyzer to rapidly detect and quantify abnormal lymphoid cells by using a restricted panel of moAbs.

Patients and Methods Seventy blood samples were analyzed by using the following moAb combinations: CD3FITC/CD4PE, CD3FITC/CD8PE in an automated assay and CD3FITC/CD19PE and CD5FITC/CD19PE by using a manual procedure which only consists of mixing EDTA blood and antibodies. Results were obtained in 4 and 15 minutes for the automated assay and for the manual procedure, respectively. The results were compared with an extensive immunophenotype performed by using the FacsCanto cytometer (Becton Dickinson).

Results Sixty out of 70 samples were evaluated:

1. the quantification of CD3/CD4 and CD3/CD8 lymphocytes was well correlated with classical flow cytometry (FC), as previously demonstrated (R² = .94 for CD3, .93 for CD3/CD4 and .95 for CD3/CD8). Interestingly, two T cell lymphomas (one with double positive T cells and one double negative T cells) were clearly recognized by the CELL DYN Sapphire and confirmed by classical FC and molecular biology.

2. B-cells immunophenotyping: 25 samples with B cell lymphoproliferative disorders and with circulating abnormal lymphocytes (20 CLL, 1 HCL, 1 B-PLL and 3 MZL) were studied. In all cases, a very good correlation between the Abbott analyzer and classical FC was observed: CD19+/CD5+ (R² = .99), CD19 (R² = .91), CD3 (R² = .91).

3. Twenty controls without abnormal circulating B lymphocytes were studied and results were similar to those obtained with classical FC (R² = .92 for CD3, .94 for CD19 and CD5). Of note in 10/70 samples, immunophenotyping using the Abbott analyzer was not possible because of the interference of lytic resistant red blood cells and/or platelets aggregates.

Conclusion The CELL-DYN Sapphire Abbott hematology analyzer provides a simple and fast immunophenotyping method useful for a preliminary screening in routine laboratory practice to detect and quantify abnormal circulating lymphoid cells simultaneously with the blood cell count and leukocyte differential.