engleski

Verification of CD34+ stem cell analysis according to ISHAGE protocol on Becton Dickinson FACSCanto

Background. Implementation of new analytical equipment into routine work requires verification procedure. We have verified flow cytometer Becton Dickinson FACSCanto II for ISHAGE protocol (Sutherland et al, 1996, J Hematotherapy, 5:213-226) stem cell (CD34+) analysis according to CLSI EP-A2-User verification of Performance for Precision and Trueness ; Approved Guideline-2nd ed., vol.25, No.17, 2005. in concordance with ISO 15189. Materials and methods. Analysis was performed on peripheral blood samples, fresh leukapheresates of peripheral blood and BC StemCell Control Kit samples. Acquisition data were analyzed using BD FACSDiva 6.0 software. Verification included assessment of random error check based upon triplicate measurement of commercial samples five days in a row, assessment of measurement uncertainty, conformation of linearity, comparison of results from reference analyzer and analysis of external quality control samples. Results. Performance characteristics: within.laboratory precision for Stem Cell Low (11, 6x10e6/L) was 9, 2%, for Stem Cell High (31, 5x10e6/L) 8, 0%. We confirmed linearity on peripheral blood sample 0 - 97, 2x10e6/L. Measurement uncertainty for Stem Cell Low was 28, 4% and Stem Cell High 19, 2%. Comparison of results with Beckman Coulter Cytomix FC500 was satisfactory (Passing Bablock, Y=-0, 01117+1, 0112x, p>0, 10). UK NEQAS for Immunophenotyping, Stem Cell enumeration scheme results were within target values (median+25 percentile) for CD34+ absolute and relative count. Conclusion. According to our verification procedure, BD FACSCanto II is cytometer of choice for performing C34+ stem cell absolute count analysis.

verification; CD34 stem cell; FACSCanto

nije evidentirano