Specimen type	Sputum or Endo-tracheal aspirate
	Optimal time of specimen collection	All specimens should be fresh and taken before antimicrobial treatment is started. Early morning freshly expectorated sputum is recommended for Mycobacterium species.
	Correct specimen type and method of collection	The material required is sputum from the lower respiratory tract expectorated by deep coughing. When the cough is dry, physiotherapy, postural drainage or inhalation of an aerosol before expectoration may be helpful. Saliva and postnasal secretions are not suitable.
	Quantity and number of specimens	Early morning specimens for examination for Mycobacterium species should be collected on 3 consecutive days.
	Laboratory analysis	Samples requesting AFB referred to TB laboratory, Liverpool for microscopy and culture
	Transport	Specimens should be transported to the laboratory as soon as possible. Sputum may be refrigerated for up to 2-3 hrs without an appreciable loss of pathogens. Any delay beyond this time may allow overgrowth of gram-negative bacilli; Haemophilus species and S. pneumoniae may die. If specimens are not processed on the same day as they are collected, results should be interpreted with care.
	Turnaround time	Negative report at 48hrs. Positive report 48-72hrs. Specimens from cystic fibrosis patients: - Negative report at 72hrs. Positive report 72hrs-6 days. B. cepacia positive report may take more than 5 days to report. Legionella spp. culture report at 10 days.

Algorithm 1: sputum for TB