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Standards for Laboratory Diagnosis of Tuberculosis. Professor Brian I. Duerden Inspector of Microbiology and Infection Control, Department of Health. TB diagnosis and management depend upon a reliable and prompt laboratory service. Guidance and Standards. National SOP How to do the tests

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Standards for Laboratory Diagnosis of Tuberculosis

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standards for laboratory diagnosis of tuberculosis

Standards for Laboratory Diagnosis of Tuberculosis

Professor Brian I. Duerden

Inspector of Microbiology and Infection Control,

Department of Health

guidance and standards
Guidance and Standards
  • National SOP
    • How to do the tests
  • NICE guidance
    • How to manage the patient
  • DH programme
    • What service should be delivered
    • 3 working groups
tb monitoring and laboratory services working group
TB monitoring and laboratory services working group
  • Surveillance standards
  • Standards for laboratory diagnosis
    • Current best practice
    • Simple and straightforward
    • Not replicate or replace the National SOP
standards to cover

Transfer to laboratory

Immediate tests


Culture, isolation and identification

Laboratory facilities and expertise


Susceptibility testing

Molecular fingerprinting/typing


PCR detection of Mtb

Immunodiagnostic tests


Standards to cover
  • Type of sample
    • Sputum (resp. sample), CSF (spinal/para-spinal/intra-cerebral), gastric washings, lymph nodes (tissues), urine, faeces
  • Number of samples
    • 2 or 3 for sputum? Consecutive days.
    • Early morning or any time?
    • True LRT specimen
  • Documentation
transfer to laboratory
Transfer to laboratory
  • Within 24h (or 1 working day, max 48h)
    • Minimise overgrowth
    • Maintain AFB character
  • Potentially infected clinical sample
    • Routine procedure
immediate tests
Immediate tests
  • Microscopy
    • Auramine fluorescent staining
    • 6-day service (not on call)
    • Perform microscopy and issue result within 24h (1 working day) of receipt
    • Telephone positive result to senior member of clinical team
    • Notify lead TB nurse, lead clinician, CCDC
  • Accreditation; IQC programme; satisfactory EQA performance; staff CPD/peer review
culture isolation and identification
Culture, isolation and identification
  • Automated liquid culture on all samples
    • Set up within 24h of receipt (6 day service)
    • Plus conventional solid culture
  • Send all isolates to RCM on day found to be positive
    • Reach RCM within 24h
  • Complete identification of most mycobacterial isolates within 21 days
identification and reporting
Identification and reporting
  • NAAT (PCR, LCR) or hybridisation gene probe for Mtb complex
    • On the day culture shows positive OR
    • Within 24h of receipt at RCM
  • Other probes and/or phenotypic tests
  • Report on day of test to
    • Senior member of clinical team
    • Lead TB nurse, lead TB clinician, CCDC
laboratory facilities and expertise
Laboratory facilities and expertise
  • Safety – Category 3 for culture
    • HSE approved
    • Contingency plan for accidental dispersal
    • Continuity plan for closure
  • Accredited
    • IQC programme, satisfactory EQA
  • Sufficient number – daily service, competence
  • Named Consultant and BMS for advice
  • Samples
    • Potentially infected samples (routine)
  • Positive cultures
    • Category A but exemption to treat as B for clinical and diagnostic purposes
      • UN 3373 – marked Diagnostic or Clinical
      • P650 packaging
      • Do not send by Royal Mail
susceptibility testing
Susceptibility testing
  • Complete within 30 days of initial receipt of clinical sample for primary agents
    • Isoniazid, rifampicin, pyrazinamide, ethambutol
      • Takes 10-20 days by liquid proportion (automated) or resistance ratio
  • Molecular detection
    • Rifampicin within 24h if MDRTB suspected
    • Isoniazid under development
  • Done at RCM with accreditation, IQC, EQA
molecular fingerprinting typing
Molecular fingerprinting/typing
    • 15-loci MIRU-VNTR
      • Mycobacterial Interspersed Repetitive Units – Variable Number Tandem Repeats
    • Results to national database
    • Other techniques as appropriate
  • Done at RCM
laboratory notification
Laboratory notification
  • HPA
    • Via CoSurv from laboratory that identifies a positive culture
    • Confirmation of positive from RCM within 24h (1 working day) of receipt
    • RCM reports culture and susceptibility results to MycobNET within 24h of report to clinician
pcr detection of mtb
PCR detection of Mtb
  • Not routine
  • Available from RCM for particular samples
    • High suspicion
    • Definitive diagnosis deemed to be urgent
    • Liaise in advance – Consultany Microbiologist to RCM
immunodiagnostic tests
Immunodiagnostic tests
  • Interferon γ (QuantiFERON-TB Gold)
  • Activated specific T-cells (T-SPOT.TB)
    • Standard under development
      • Which patients?
      • How long should it take?
      • Who provides it?
      • What do the results mean and who interprets them?
  • Report within 3 days of receipt
  • Inform the Microbiology service
    • Ensure same reporting as for positive microscopy and culture results
  • Send autopsy samples to Microbiology without formalin!!
  • [Role of PCR to be determined]
implementation of standards
Implementation of standards
  • Local responsibility
    • What is done where?
      • Microscopy; culture; identification
    • What throughput is needed?
    • Equipment – cost-effectiveness
    • Personnel
      • Maintain skills; CPD; peer review
      • Named individuals for advice
      • Back-up and cover
    • IQC, EQA
standards for quality
Standards for Quality

Only do what you can do properly!