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Reliability of a rapid hematology stain for sputum cytology.

Gonçalves J, Pizzichini E, Pizzichini MM, Steidle LJ, Rocha CC, Ferreira SC, Zimmermann CT - J Bras Pneumol (2014 May-Jun)

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

View Article: PubMed Central - PubMed

Affiliation: Clinical Analysis Department, University Hospital, Federal University of Santa Catarina, Florianópolis, Brazil.

ABSTRACT

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.

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Related in: MedlinePlus

Bland & Altman plots. Interobserver reproducibility for the proportionof neutrophils (in A), eosinophils (in B), and macrophages (in C) on thecytospin slides prepared from induced sputum samples and stained withMay-Grünwald-Giemsa (MGG). Intraobserver reproducibility for the proportion ofneutrophils (in D), eosinophils (in E), and macrophages (in F) on the cytospinslides prepared from induced sputum samples and stained by either of the twostaining techniques. The plots refer to the differences between the readings byobservers 1 and 2 (y axis) in relation to the mean readings by observers 1 and2 (x axis). The central broken line indicates absence of differences, and theperipheral broken lines indicate two standard deviations of the mean of thedifferences. ICC: intraclass correlation coefficient; NEU.: neutrophils; EOS.:eosinophils; and MAC.: macrophages.
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f03: Bland & Altman plots. Interobserver reproducibility for the proportionof neutrophils (in A), eosinophils (in B), and macrophages (in C) on thecytospin slides prepared from induced sputum samples and stained withMay-Grünwald-Giemsa (MGG). Intraobserver reproducibility for the proportion ofneutrophils (in D), eosinophils (in E), and macrophages (in F) on the cytospinslides prepared from induced sputum samples and stained by either of the twostaining techniques. The plots refer to the differences between the readings byobservers 1 and 2 (y axis) in relation to the mean readings by observers 1 and2 (x axis). The central broken line indicates absence of differences, and theperipheral broken lines indicate two standard deviations of the mean of thedifferences. ICC: intraclass correlation coefficient; NEU.: neutrophils; EOS.:eosinophils; and MAC.: macrophages.

Mentions: Regarding intraobserver agreement, the ICC values for the two observers for thedifferential cell counts on the pairs of cytospin slides stained by either of the twostudied techniques indicated that it was almost perfect for neutrophils (ICC = 0.97 forboth), eosinophils (ICC = 0.99 and 0.98), and macrophages (ICC = 0.96 for both). Forlymphocyte counts, intraobserver agreement was substantial for observer 1 (ICC = 0.75)and moderate for observer 2 (ICC = 0.65). These results are shown graphically in Figure 3.(17)


Reliability of a rapid hematology stain for sputum cytology.

Gonçalves J, Pizzichini E, Pizzichini MM, Steidle LJ, Rocha CC, Ferreira SC, Zimmermann CT - J Bras Pneumol (2014 May-Jun)

Bland & Altman plots. Interobserver reproducibility for the proportionof neutrophils (in A), eosinophils (in B), and macrophages (in C) on thecytospin slides prepared from induced sputum samples and stained withMay-Grünwald-Giemsa (MGG). Intraobserver reproducibility for the proportion ofneutrophils (in D), eosinophils (in E), and macrophages (in F) on the cytospinslides prepared from induced sputum samples and stained by either of the twostaining techniques. The plots refer to the differences between the readings byobservers 1 and 2 (y axis) in relation to the mean readings by observers 1 and2 (x axis). The central broken line indicates absence of differences, and theperipheral broken lines indicate two standard deviations of the mean of thedifferences. ICC: intraclass correlation coefficient; NEU.: neutrophils; EOS.:eosinophils; and MAC.: macrophages.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4109197&req=5

f03: Bland & Altman plots. Interobserver reproducibility for the proportionof neutrophils (in A), eosinophils (in B), and macrophages (in C) on thecytospin slides prepared from induced sputum samples and stained withMay-Grünwald-Giemsa (MGG). Intraobserver reproducibility for the proportion ofneutrophils (in D), eosinophils (in E), and macrophages (in F) on the cytospinslides prepared from induced sputum samples and stained by either of the twostaining techniques. The plots refer to the differences between the readings byobservers 1 and 2 (y axis) in relation to the mean readings by observers 1 and2 (x axis). The central broken line indicates absence of differences, and theperipheral broken lines indicate two standard deviations of the mean of thedifferences. ICC: intraclass correlation coefficient; NEU.: neutrophils; EOS.:eosinophils; and MAC.: macrophages.
Mentions: Regarding intraobserver agreement, the ICC values for the two observers for thedifferential cell counts on the pairs of cytospin slides stained by either of the twostudied techniques indicated that it was almost perfect for neutrophils (ICC = 0.97 forboth), eosinophils (ICC = 0.99 and 0.98), and macrophages (ICC = 0.96 for both). Forlymphocyte counts, intraobserver agreement was substantial for observer 1 (ICC = 0.75)and moderate for observer 2 (ICC = 0.65). These results are shown graphically in Figure 3.(17)

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

View Article: PubMed Central - PubMed

Affiliation: Clinical Analysis Department, University Hospital, Federal University of Santa Catarina, Florianópolis, Brazil.

ABSTRACT

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.

Show MeSH
Related in: MedlinePlus