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A new assay for quantifying human reticulocytes.

E R Burns, S N Goldberg, B Wenz

    American Journal of Clinical Pathology
    |September 1, 1987
    PubMed
    Summary
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    This study introduces a novel, precise method for quantitative reticulocyte analysis using standard blood cell analyzers and density gradient centrifugation. The new assay significantly improves accuracy and precision over manual reticulocyte counts.

    Area of Science:

    • Hematology
    • Clinical Pathology
    • Biomedical Engineering

    Background:

    • Accurate reticulocyte counting is crucial for diagnosing and monitoring various anemias and bone marrow disorders.
    • Conventional microscopic reticulocyte counts are labor-intensive and suffer from significant inter-observer variability and limited precision.
    • There is a need for a more accurate, precise, and automated method for reticulocyte quantification.

    Purpose of the Study:

    • To develop and validate a new quantitative reticulocyte analysis method.
    • To improve the accuracy and precision of reticulocyte enumeration compared to existing techniques.
    • To utilize conventional clinical blood cell analyzers for enhanced reticulocyte analysis.

    Main Methods:

    • A novel assay involving density gradient centrifugation of blood using Stractan (arabino-galactan polysaccharide).

    Related Experiment Videos

  • Enrichment of neocytes (young red blood cells) in the Stractan layer via centrifugation (1,500 x g for 30 minutes).
  • Measurement of mean corpuscular volume (MCV) of red blood cells in the Stractan layer and whole blood, with reticulocyte percentage determined by MCV ratio and standard curve.

    • Main Results:

    • The method demonstrated excellent linearity and correlation (r = 0.96) with manual reticulocyte counts in 93 patients.
    • High agreement (within 0.2%) was observed for single sample results.
    • The assay exhibited superior precision, with within-run and between-run coefficients of variation less than 1%.

    Conclusions:

    • The described assay provides a highly accurate and precise method for quantitative reticulocyte analysis.
    • This technique leverages conventional hematology analyzers, offering a significant improvement (at least 20-fold) in accuracy and precision over manual methods.
    • The Stractan density gradient method offers a robust and reproducible approach for reticulocyte enumeration in clinical settings.