title: A method to preserve sputum for delayed examination.
abstract: Examination of sputum cell counts is limited by the need to process samples within hours of expectoration. The validity and repeatability of a method to preserve sputum for delayed processing and examination were investigated in this study. Portions of selected sputum from 39 subjects were dispersed with dithiothreitol (routine method), or were fixed in a dithiothreitol-formaldehyde mixture for 48-72 h before dispersal with trypsin (preservation method). Total and differential cell counts using the two methods were compared; within-method repeatability of the preservation method was also examined. The intraclass correlation coefficient (ICC) for total cell counts and percentage of eosinophils, neutrophils and macrophages in sputum processed by the two methods was 0.89, 0.86, 0.91 and 0.90, respectively. Within-method repeatability (ICC) of the preservation method for the same cellular indices was 0.99, 0.94, 0.97 and 0.97, respectively. The interobserver repeatability for eosinophils, neutrophils and macrophages was 0.96, 0.97 and 0.97 using the preservation method, and 0.96, 0.99 and 0.99 using the routine method, respectively. This method of sputum preservation and dispersal is valid, reliable and convenient, and may be used for delayed processing and examination.
authors.affiliation: Asthma Research Group, Firestone Institute for Respiratory Health, St Joseph's Healthcare-McMaster University, Hamilton, ON, Canada.
authors.forename: M M
authors.initials: MM
authors.lastname: Kelly
authors.forename: F E
authors.initials: FE
authors.lastname: Hargreave
authors.forename: G
authors.initials: G
authors.lastname: Cox
mesh_terms: D002452:Cell Count / Q000379:methods*; D003430:Cross-Sectional Studies; D004229:Dithiothreitol / Q000494:pharmacology; D005260:Female; D005404:Fixatives / Q000494:pharmacology; D005557:Formaldehyde / Q000494:pharmacology; D006801:Humans; D007202:Indicators and Reagents / Q000494:pharmacology; D008297:Male; D008875:Middle Aged; D011446:Prospective Studies; D015203:Reproducibility of Results; D016037:Single-Blind Method; D013048:Specimen Handling / Q000379:methods*; D013183:Sputum / Q000166:cytology* / Q000187:drug effects*; D013997:Time Factors; D014357:Trypsin / Q000494:pharmacology
chemical_list: D005404:Fixatives; D007202:Indicators and Reagents; D005557:Formaldehyde; D014357:Trypsin; D004229:Dithiothreitol