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Comparison of sputum processing techniques in cystic fibrosis
1Discipline of Paediatrics, University of Newcastle, New South Wales, Australia.
Pediatric Pulmonology
|December 1, 1996
Summary
Sputum analysis for cystic fibrosis (CF) requires an enzyme mixture (E) for accurate cell counts, as dithiothreitol (DTT) processing, used in asthma studies, yields false results in CF sputum. This enzyme method improves total and neutrophil cell counts without affecting viability.
Area of Science:
- Pulmonary Medicine
- Biochemistry
- Cell Biology
Background:
- Sputum analysis is crucial for studying airway inflammation.
- Dithiothreitol (DTT) is standard for sputum processing in asthma.
- Its effectiveness in cystic fibrosis (CF) sputum is unestablished.
Purpose of the Study:
- Compare DTT with an enzyme mixture (E) for CF sputum processing.
- Evaluate sample quality, cell viability, total cell count (TCC), neutrophil count, and elastase immunoreactivity.
Main Methods:
- Sputum from 11 CF patients was split and processed with DTT or enzyme mixture (E).
- Assessed outcomes included sample quality, cell viability, TCC, neutrophil count, and elastase immunoreactivity.
- Purified neutrophils were incubated with DTT and E to assess toxicity.
Main Results:
- Sample quality and cell viability were similar between DTT and E methods.
- Enzyme mixture (E) yielded significantly higher TCC and neutrophil counts (P=0.01, P=0.02).
- Elastase immunoreactivity was lost with E processing (P=0.04); DTT showed higher retention.
Conclusions:
- Standard asthma sputum processing with DTT is unsuitable for CF.
- An enzyme mixture (E) provides more accurate cell counts for CF sputum.
- Incomplete dispersal, not toxicity, explains lower neutrophil counts with DTT in CF sputum.