Professional Documents
Culture Documents
LABORATORY
INVESTIGATIONS
by
Dr. Zubaidah Abdul Wahab
Datin Dr. Ganeswrie Raj
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LEARNING OBJECTIVES
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INTRODUCTION
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INTRODUCTION
• Specimen collection
• Lab investigations
– Microscopy
– Culture
– Identification
– AntiTB sensitivity
– Molecular
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SPUTUM COLLECTION
• Sputum1
• At least 2 specimens
• At least one early morning specimen
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OTHER SAMPLES
• Sterile body fluids
• include blood, CSF, pleural fluid, peritoneal
fluid
• should be collected aseptically
• transported immediately to laboratory in
sterile container
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MICROSCOPY
• Microscopy
• Presumptive diagnosis
• Sputum
• Ziehl-Neelsen staining for AFB
• Conventional microscope
• low sensitivity (20 - 60%)1
• Light emitting diode-based
fluorescence microscopy (LED FM)2
• 10% more sensitive
• shorter time spent
• quicker turnaround time
1Steingart KR et al., Lancet Infect Dis, 2006
2Shenai S et al., Int J Tuberc Lung Dis, 2011
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CONVENTIONAL LIGHT MICROSCOPY
• Culture
• conventional method using LJ or Ogawa
• 6 - 8 weeks
• Drug-susceptibility testing
• 1 - 2 months
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PHENOTYPIC METHOD –
Proportion method
A B
M. tuberculosis Susceptible
to INH
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NUCLEIC ACID
AMPLIFICATION TEST (NAAT)
• NAAT
• molecular technique
• provide rapid results within 24 - 48 hours
• ability to confirm presence of Mycobacterium in 50 -
80% AFB smear negative & culture positive1,2
• detect Mycobacterium in specimens weeks earlier
than culture for 80 - 90% patients2
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LINE PROBE ASSAY (LPA)
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GENE XPERT MTB
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COMMERCIAL SEROLOGICAL ASSAY
• Recommendation 5
Commercial serological assay should not
be used to diagnose pulmonary &
extrapulmonary TB.
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COMMERCIAL SEROLOGICAL ASSAY
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ROLE OF ADENOSINE DEAMINASE (ADA)
IN TB DIAGNOSIS
• ADA is an enzyme involved in purine
metabolism of the cells.
• Measurement of ADA level in pleural effusion
& CSF is useful additional test in pleural &
meningeal TB.
• In pleural TB, sensitivity = 92% & specificity = 90%1
• In CSF-ADA in TB meningitis, sensitivity = 93% &
specificity of 97% if cut-off level of >10 U/L used2
1Liang QL et al., Respir Med, 2008
2Rana SV et al., Clin Neurol Neurosurg., 2010
TAKE HOME MESSAGES
dr_zubaidah@sel.moh.gov.my
ganiraj_balan@yahoo.com
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