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Laboratory and Animal Investigations |

Comparison of Three Methods for Differential Cell Count in Induced Sputum*

Beatriz M. Saraiva-Romanholo; Viviani Barnabé; Ana Lúcia I. Carvalho; Milton A. Martins; Paulo H. N. Saldiva; Maria do Patrocínio T. Nunes
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*From the Departments of Medicine (Mss. Saraiva-Romanholo and Barnabé, and Drs. Carvalho and Martins) and Pathology (Drs. Saldiva and Nunes), School of Medicine, University of São Paulo, São Paulo, Brazil.

Correspondence to: Maria do Patrocínio Tenório Nunes, MD, PhD, Av. Dr Arnaldo, 455 sala 1216, São Paulo, SP, Brazil, 01246-903; e-mail: ppatro@usp.br



Chest. 2003;124(3):1060-1066. doi:10.1378/chest.124.3.1060
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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.

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