Comparison of three methods for differential cell count in induced sputum

Chest. 2003 Sep;124(3):1060-6. doi: 10.1378/chest.124.3.1060.


Background: Induced sputum (IS) using the cytospin technique has been extensively employed to characterize inflammatory airway diseases; however, procedures of cell enrichment based on cytospin increase the analytical costs and require slide processing within a short period of time after sampling.

Study objectives: To compare three different techniques for cytologic analysis of IS, and to determine the time required by each method and the costs involved.

Design: Cross-sectional study.

Setting: Tertiary-care university hospital.

Patients and measurements: Eighty-nine patients with asthma and 11 subjects without asthma were submitted to increasing hypertonic saline solution concentrations of 2, 3, 4, and 5% for 7 min for sputum induction. Samples were smeared without treatment with 0.1% dithiothreitol (DTT) [technique A], after treatment with DTT (technique B), and after treatment with DTT and cytospin (technique C). All slides were air-dried and stained with Leishman stain. Two independent observers counted at least 200 inflammatory cells on each slide.

Results: Eighty percent of the slides processed by techniques A and B and 65% of the slides processed by technique C represented sputum samples of acceptable quality. The eosinophil percentages in sputum obtained by techniques A and C were closely correlated, as also were those obtained by techniques B and C (r = 0.64 and r = 0.63, respectively; p < 0.01). There was a positive correlation for eosinophils when we compared techniques A and B (r = 0.57, p < 0.01). The neutrophil correlation was significant when the three techniques were compared (technique A vs technique B, r = 0.66; technique A vs technique C, r = 0.51; and technique B vs technique C, r = 0.57; p < 0.01). Bland-Altman analysis showed a good agreement for eosinophil and neutrophil counts when techniques A and B were compared to technique C.

Conclusions: The three techniques are good indicators of lung inflammation. Techniques A and B are less time consuming and are of lower cost.

Publication types

  • Comparative Study
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Adolescent
  • Adult
  • Aged
  • Asthma / diagnosis
  • Asthma / immunology*
  • Child
  • Cross-Sectional Studies
  • Female
  • Humans
  • Leukocyte Count / methods*
  • Lymphocyte Count / methods*
  • Male
  • Middle Aged
  • Peak Expiratory Flow Rate / physiology
  • Reference Values
  • Sensitivity and Specificity
  • Sputum / cytology*
  • Sputum / immunology