Suitable clinical samples should ideally be sent for conventional AFB microscopy and culture investigations as outlined above. Note: It should be noted that if there is not enough specimen volume for PCR and culture, then only culture should be done. All samples, even if PCR positive, should be submitted for culture. 4.5.5 Culture media, conditions and organisms Clinical details/ conditions Specimen Standard media Incubation Cultures read Target organism(s) ‡ Temp oC Atmos Time All conditions Sterile samples: All bone marrow aspirates Wound swab Pus or Pus swabs* CSF Ascites Joint fluid Pleural fluid Automated liquid systems If automated monitoring systems are used, after decontamination of clinical samples, refer to local protocols and manufacturer’s recommendations. These should be checked continuously for bottles that flag positive. A blood agar plate should be set up for all bottles that are flagged positive to check for contamination. Mycobacterium species LJ + pyruvate 35 – 37 Air 8 weeks extending to 12 weeks if required58 Weekly Non-sterile samples: Sputum washings Gastric washings Automated liquid systems If automated monitoring systems are used, after decontamination of clinical samples, refer to local protocols and manufacturer’s recommendations. These should be checked continuously for bottles that flag positive. A blood agar plate should be set up for all bottles that are flagged positive to check for contamination. Mycobacterium tuberculosis LJ + pyruvate** 35 – 37 Air 8 weeks extending to 12 weeks if required58 Weekly Genitourinary tuberculosis Urine Automated liquid systems If automated monitoring systems are used, after decontamination of clinical samples, refer to local protocols and manufacturer’s recommendations. These should be checked continuously for bottles that flag positive. A blood agar plate should be set up for all bottles that are flagged positive to check for contamination. Mycobacterium species LJ + pyruvate 35 – 37 Air 8 weeks extending to 12 weeks if required58 Weekly
Skin infections - ‘fish tank granuloma’ - cutaneous ulcer (or “Buruli” or “Bairnsdale ulcer” Skin lesions Tissue Biopsy LJ + pyruvate 28 - 30 Air 8 weeks extending to 12 weeks if required58 Weekly M. marinum M. ulcerans M. chelonae Cardiac surgery Post-surgery sites eg sternotomy Automated liquid systems If automated monitoring systems are used, after decontamination of clinical samples, refer to local protocols and manufacturer’s recommendations. These should be checked continuously for bottles that flag positive. A blood agar plate should be set up for all bottles that are flagged positive to check for contamination. M.chimaera / M. avium – intracellulare group LJ + pyruvate 35 - 37 Air 8 weeks extending to 12 weeks if required58 Weekly For these situations, add the following: Clinical details/ Conditions Specimen Supplementary media Incubation Cultures read Target organism(s) Temp oC Time Tuberculous meningitis CSF and other non-sputum specimens Kirchner’s liquid media OR Middlebrook 7H9 35 – 37 Air 8 weeks extending to 12 weeks if required58 Weekly Mycobacterium species Clinical details/ Conditions Specimen Optional media Incubation Cultures read Target organism(s) Temp oC Atmos Time Osteomyelitis Consider for bone and fluids eg from extremities and/or direct smear positive Automated liquid systems* If automated monitoring systems are used, after decontamination of clinical samples, refer to local protocols and manufacturer’s recommendations. These should be checked continuously for bottles that flag positive. A blood agar plate should be set up for all bottles that are flagged positive to check for contamination. Non-tuberculous mycobacteria LJ + pyruvate 28 - 30 Air 8 weeks extending to 12 weeks if required58 Weekly Pulmonary tuberculosis Smear +ve BAL/washing LJ + pyruvate 28 – 30 Air 4-6 weeks Weekly Rapidly growing mycobacteria Pulmonary tuberculosis Sputum Brochoalveolar washings Gastric washings LJ + glycerol Note: M.xenopi do not grow well on medium with pyruvate preferring glycerol. 42 Air 8 weeks extending to 12 weeks if required58 Weekly M. xenopi