title: Automated reticulocyte counting and immature reticulocyte fraction measurement. Comparison of ABX PENTRA 120 Retic, Sysmex R-2000, flow cytometry, and manual counts.
abstract: We evaluated reticulocyte counting and measurement of immature reticulocyte fraction (IRF) with the ABX PENTRA 120 Retic blood analyzer on 300 blood samples. Reticulocyte counts were compared with those obtained by visual counting of 2,000 RBCs, by the TOA (Kobe, Japan) Sysmex R-2000 and a flow cytometry method. The parameters analyzed were the percentages of reticulocytes on all analyzers and the IRF with different modalities. The Retic Count kit (Becton Dickinson, San Jose, CA) was used with the Coulter (Hialech, FL) XL, and a mean channel of fluorescence (MCF) was calculated to fit the reticulocyte maturation. Reticulocyte counting with the ABX (Montpellier, France) PENTRA 120 Retic showed excellent precision and linearity with no significant carryover. Reticulocyte counts were stable after blood storage for 72 hours at 4 degrees C but not at room temperature (RT). IRF parameters values were stable for only 8 hours at 4 degrees C and 6 hours at RT. Comparisons of the methods showed good intraclass correlation (RI) for reticulocyte percentages between ABX PENTRA 120 Retic and Sysmex R-2000, ABX PENTRA 120 Retic and flow cytometry, Sysmex R-2000 and flow cytometry, and ABX PENTRA 120 Retic and manual counting. IRF values were correlated between fluorescence rates and RNA content, but in each case, low RI values were found, showing that Sysmex and ABX IRF values were not concordant. We obtained a significant correlation between mean fluorescence index and the MCF measured by flow cytometry, but the 2 methods were not concordant using the RI. The ABX PENTRA 120 Retic is a good instrument for analyzing reticulocyte count and percentage and allows a good analysis of IRF with several modalities.
authors.affiliation: Laboratory of Hematology, Hôpital Haut-Lévèque, CHU Bordeaux, France.
authors.forename: F
authors.initials: F
authors.lastname: Lacombe
authors.forename: L
authors.initials: L
authors.lastname: Lacoste
authors.forename: J P
authors.initials: JP
authors.lastname: Vial
authors.forename: A
authors.initials: A
authors.lastname: Briais
authors.forename: J
authors.initials: J
authors.lastname: Reiffers
authors.forename: M R
authors.initials: MR
authors.lastname: Boisseau
authors.forename: P
authors.initials: P
authors.lastname: Bernard
mesh_terms: D000208:Acute Disease; D001331:Automation; D002908:Chronic Disease; D005069:Evaluation Studies as Topic; D005434:Flow Cytometry / Q000379:methods; D005456:Fluorescent Dyes; D006801:Humans; D007938:Leukemia / Q000097:blood; D015203:Reproducibility of Results; D017701:Reticulocyte Count / Q000295:instrumentation / Q000379:methods*; D012156:Reticulocytes / Q000166:cytology*; D012680:Sensitivity and Specificity
chemical_list: D005456:Fluorescent Dyes