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Comparative Study
. 2014 May-Jun;40(3):250-8.
doi: 10.1590/s1806-37132014000300008.

Reliability of a rapid hematology stain for sputum cytology

[Article in English, Portuguese]
Jéssica Gonçalves  1 , Emilio Pizzichini  2 , Marcia Margaret Menezes Pizzichini  2 , Leila John Marques Steidle  3 , Cristiane Cinara Rocha  4 , Samira Cardoso Ferreira  1 , Célia Tânia Zimmermann  5
Affiliations
Comparative Study

Reliability of a rapid hematology stain for sputum cytology

[Article in English, Portuguese]
Jéssica Gonçalves et al. J Bras Pneumol. 2014 May-Jun.

Abstract
in English, Portuguese

Objective: To determine the reliability of a rapid hematology stain for the cytological analysis of induced sputum samples.

Methods: This was a cross-sectional study comparing the standard technique (May-Grünwald-Giemsa stain) with a rapid hematology stain (Diff-Quik). Of the 50 subjects included in the study, 21 had asthma, 19 had COPD, and 10 were healthy (controls). From the induced sputum samples collected, we prepared four slides: two were stained with May-Grünwald-Giemsa, and two were stained with Diff-Quik. The slides were read independently by two trained researchers blinded to the identification of the slides. The reliability for cell counting using the two techniques was evaluated by determining the intraclass correlation coefficients (ICCs) for intraobserver and interobserver agreement. Agreement in the identification of neutrophilic and eosinophilic sputum between the observers and between the stains was evaluated with kappa statistics.

Results: In our comparison of the two staining techniques, the ICCs indicated almost perfect interobserver agreement for neutrophil, eosinophil, and macrophage counts (ICC: 0.98-1.00), as well as substantial agreement for lymphocyte counts (ICC: 0.76-0.83). Intraobserver agreement was almost perfect for neutrophil, eosinophil, and macrophage counts (ICC: 0.96-0.99), whereas it was moderate to substantial for lymphocyte counts (ICC = 0.65 and 0.75 for the two observers, respectively). Interobserver agreement for the identification of eosinophilic and neutrophilic sputum using the two techniques ranged from substantial to almost perfect (kappa range: 0.91-1.00).

Conclusions: The use of Diff-Quik can be considered a reliable alternative for the processing of sputum samples.

Objetivo:: Determinar a confiabilidade da coloração hematológica rápida para a análise do escarro induzido.

Métodos:: Estudo transversal comparando a técnica padrão (coloração May-Grünwald-Giemsa) com a coloração hematológica rápida (panótico rápido). Participaram do estudo 50 indivíduos (21 asmáticos, 19 portadores de DPOC e 10 controles). Após a coleta do escarro induzido, foram preparadas 4 lâminas, sendo 2 coradas por May-Grünwald-Giemsa e 2 por panótico rápido. As lâminas foram lidas de forma independente por dois pesquisadores capacitados para o exame de escarro induzido e cegados para a identificação das lâminas. A confiabilidade para as contagens celulares dos dois métodos foi avaliada pela determinação dos coeficientes de correlação intraclasse (CCI) para as concordâncias intraobservador e interobservador. As concordâncias na identificação de escarro neutrofílico e eosinofílico entre observadores e entre as duas colorações foram calculadas por estatística kappa.

Resultados:: Nas duas colorações, os CCI apontaram concordância interobservador quase perfeita para as contagens de neutrófilos, eosinófilos e macrófagos (variação do CCI: 0,98-1,00) e substancial para as contagens de linfócitos (variação do CCI: 0,76-0,83). Na análise intraobservador, a concordância foi quase perfeita para as contagens de neutrófilos, eosinófilos e macrófagos (variação do CCI: 0,96-0,99) e de moderada a substancial para as contagens de linfócitos (CCI = 0,65 e 0,75 para observadores 1 e 2, respectivamente). A concordância interobservador na identificação de escarro eosinofílico e neutrofílico para os dois métodos de coloração variou entre substancial e quase perfeita (variação kappa: 0,91-1,00).

Conclusões:: O panótico rápido pode ser considerado uma alternativa confiável para o processamento de amostras de escarro.

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Figures

Figure 1
Figure 1. Induced sputum differential cell counts in the three groups studied.
Figure 2
Figure 2. Median differential cell counts on the cytospin slides stained either with May-Grünwald-Giemsa (dark gray bar) or Diff-Quik (light gray bar)
Figure 3
Figure 3. Bland & Altman plots. Interobserver reproducibility for the proportion of neutrophils (in A), eosinophils (in B), and macrophages (in C) on the cytospin slides prepared from induced sputum samples and stained with May-Grünwald-Giemsa (MGG). Intraobserver reproducibility for the proportion of neutrophils (in D), eosinophils (in E), and macrophages (in F) on the cytospin slides prepared from induced sputum samples and stained by either of the two staining techniques. The plots refer to the differences between the readings by observers 1 and 2 (y axis) in relation to the mean readings by observers 1 and 2 (x axis). The central broken line indicates absence of differences, and the peripheral broken lines indicate two standard deviations of the mean of the differences. ICC: intraclass correlation coefficient; NEU.: neutrophils; EOS.: eosinophils; and MAC.: macrophages.
Figura 1
Figura 1. Celularidade diferencial no escarro induzido nos três grupos do estudo.
Figura 2
Figura 2. Mediana da contagem celular diferencial em lâminas coradas com May-Grünwald-Giemsa (barra cinza escuro) e pela coloração hematológica rápida (barra cinza claro).
Figura 3
Figura 3. Representações gráficas de Bland & Altman. Reprodutibilidade interobservador das proporções de neutrófilos (em A), eosinófilos (em B) e macrófagos (em C) no escarro induzido em lâminas coradas por May-Grünwald-Giemsa (MGG). Reprodutibilidade intraobservador das proporções de neutrófilos (em D), eosinófilos (em E) e macrófagos (em F) no escarro induzido em lâminas coradas pelos dois métodos de coloração. As representações se referem às diferenças entre as leituras dos observadores 1 e 2 (eixo y) em relação à média das leituras dos observadores 1 e 2 (eixo x). A linha interrompida central indica ausência de diferenças e as linhas interrompidas periféricas indicam dois desvios-padrão da média das diferenças. CCI: coeficiente de correlação intraclasse; NEU.: neutrófilos; EOS.: eosinófilos; e MAC.: macrófagos.
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References

    1. Djukanovic R, Sterk PJ, editors. An atlas of induced sputum--an aid for research and diagnosis. New York: Parthenon Publishing Group; 2004.
    1. Pizzichini E, Efthimiadis A, Pizzichini MM, Hargreave FE. Sputum Examination for indices of airway inflammation: laboratory procedures. Lund, Sweden: Astra Draco AB; 1997.
    1. Silva LC, Hetzel JL. Pneumonias obstrutivas. In: Silva LC, editor. Condutas em Pneumologia. Rio de Janeiro: Revinter; 2001. pp. 261–352.
    1. Pavord ID, Sterk PJ, Hargreave FE, Kips JC, Inman MD, Louis R, et al. Clinical applications of assessment of airway inflammation using induced sputum. Eur Respir J Suppl. 2002;37:40s–43s. - PubMed
    1. Pavord ID, Brightling CE, Woltmann G, Wardlaw AJ. Non-eosinophilic corticosteroid unresponsive asthma. Lancet. 1999;353(9171):2213–2214. http://dx.doi.org/10.1016/S0140-6736(99)01813-9 - DOI - PubMed

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