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Related Experiment Video

Updated: Jun 14, 2026

Methodology for Sputum Induction and Laboratory Processing

Methodology for Sputum Induction and Laboratory Processing

Published on: December 17, 2017

Do processing time and storage of sputum influence quantitative bacteriology in bronchiectasis?

Maeve P Murray¹, Catherine J Doherty², John R W Govan²

¹Department of Respiratory Medicine, Royal Infirmary of Edinburgh, 51 Little France Crescent, Old Dalkeith Road, Edinburgh EH16 4SA, UK.

Journal of Medical Microbiology

|March 27, 2010

Summary

Sputum bacterial density is stable for up to 6 hours at room temperature. Refrigerated storage (4°C) is recommended for longer periods, while freezing (-20°C) reduces bacterial counts.

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Area of Science:

Microbiology
Clinical Diagnostics
Respiratory Medicine

Background:

Accurate bacterial quantification in sputum is crucial for diagnosing and managing respiratory infections like bronchiectasis.
Spontaneous sputum collection is a common method, but sample handling conditions can impact bacterial viability and density.

Purpose of the Study:

To investigate the effect of storage time and temperature on the bacterial density of spontaneous sputum samples.
To determine optimal storage conditions for maintaining the integrity of bacterial populations in sputum.

Main Methods:

Sputum samples were collected from ten bronchiectasis patients over 45 minutes.
Aliquots were processed immediately (1-6 hours at 25°C) or stored at 25°C, 4°C, or -20°C for 24 and 48 hours before processing.
Quantitative bacteriology (colony-forming units per milliliter) was performed for Pseudomonas aeruginosa and Staphylococcus aureus.

Main Results:

Pseudomonas aeruginosa density remained stable in samples processed up to 6 hours post-expectoration at 25°C.
Storage at 4°C for 24-48 hours did not significantly alter bacterial load compared to immediate processing.
Storage at -20°C significantly reduced bacterial density, while storage at 25°C for 24-48 hours significantly increased it.

Conclusions:

Sputum bacterial density is unaffected by storage for up to 6 hours at 25°C.
Refrigeration at 4°C is a suitable method for preserving sputum bacterial load for up to 48 hours.
Freezing at -20°C or prolonged room temperature storage should be avoided due to significant alterations in bacterial density.