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AFB-DNA (TB-PCR) Detection by RTPCR, Reflex to Rifampicin resistance by Ultra CBNAAT (Specimen)

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Same as above but applied to any specimen type.

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AFB-DNA (TB-PCR) Detection by RTPCR Reflex to Rifampicin resistance by Ultra CBNAAT

  • Why is it done?
    • Detection of Mycobacterium tuberculosis (MTB) DNA using Real-Time PCR technology to confirm active tuberculosis infection
    • Simultaneous identification of rifampicin resistance mutations using Ultra CBNAAT (Cartridge Based Nucleic Acid Amplification Test) as a reflex test
    • Rapid diagnosis of TB in patients with respiratory symptoms (cough, fever, night sweats, weight loss)
    • Early detection of drug-resistant tuberculosis (MDR-TB) to guide appropriate treatment protocols
    • Confirmation of TB diagnosis in patients with positive sputum smear microscopy or clinical suspicion
    • Initial diagnostic testing for patients with suspected TB, including those with HIV co-infection
    • Monitoring treatment response and detection of treatment failure or relapse
  • Normal Range
    • MTB DNA Detection: NEGATIVE (NOT DETECTED) - indicates no tuberculosis DNA detected in the specimen
    • Rifampicin Resistance: NOT DETECTED - indicates susceptibility to rifampicin (drug-sensitive TB)
    • Interpretation of Results:
    • NEGATIVE: No MTB DNA detected - patient does not have active TB infection; normal finding
    • POSITIVE: MTB DNA detected with Rifampicin SUSCEPTIBLE - patient has drug-sensitive TB
    • POSITIVE: MTB DNA detected with Rifampicin RESISTANT - patient has MDR-TB (Multi-Drug Resistant TB)
    • INCONCLUSIVE/ERROR: Test failed to produce clear results; specimen may require re-collection or repeat testing
  • Interpretation
    • MTB DNA NEGATIVE:
    • No evidence of active tuberculosis infection
    • Rules out TB as the cause of respiratory symptoms; consider alternative diagnoses
    • Note: Negative result does not exclude TB if clinical suspicion remains high; repeat testing may be warranted
    • MTB DNA POSITIVE with Rifampicin SUSCEPTIBLE:
    • Confirms diagnosis of active tuberculosis
    • Indicates susceptibility to rifampicin - patient can be treated with standard first-line anti-TB drugs
    • Standard TB regimen recommended (Isoniazid, Rifampicin, Pyrazinamide, Ethambutol)
    • MTB DNA POSITIVE with Rifampicin RESISTANT:
    • Confirms diagnosis of Multi-Drug Resistant TB (MDR-TB) - resistance to both isoniazid and rifampicin
    • Requires modification of treatment regimen to include second-line anti-TB drugs
    • Treatment typically includes fluoroquinolones, injectable agents, and bedaquiline or linezolid
    • Associated with longer treatment duration and higher risk of treatment failure
    • Factors Affecting Test Accuracy:
    • Specimen quality and type (sputum, bronchoalveolar lavage, cerebrospinal fluid)
    • Bacterial load in specimen - higher bacillary load increases sensitivity
    • Stage of TB infection - sensitivity lower in early disease
    • HIV co-infection status - may affect bacillary load and test performance
    • Presence of inhibitors in specimen that may affect PCR amplification
  • Associated Organs
    • Primary Organ Systems:
    • Respiratory System (Lungs) - primary site of infection in pulmonary TB
    • Immune System - TB affects macrophages and CD4+ T cells
    • Medical Conditions Associated with Abnormal Results:
    • Pulmonary Tuberculosis (PTB) - most common form affecting lungs
    • Extrapulmonary Tuberculosis (EPTB) - TB affecting other organs including lymph nodes, pleura, meninges, bones
    • Multi-Drug Resistant TB (MDR-TB) - TB resistant to isoniazid and rifampicin
    • Extensively Drug-Resistant TB (XDR-TB) - resistance to fluoroquinolones and injectable drugs
    • TB-HIV co-infection - patients with HIV have higher TB risk
    • Potential Complications of Abnormal Results:
    • Cavitary lung disease with potential hemoptysis
    • Progressive respiratory failure if left untreated
    • Disseminated TB with involvement of multiple organs
    • TB meningitis - serious neurological complication with high mortality
    • MDR-TB complications - treatment resistance and higher mortality rates
    • Secondary bacterial infections and bronchiectasis
  • Follow-up Tests
    • If MTB DNA POSITIVE with Susceptibility (Drug-Sensitive TB):
    • Chest X-ray - evaluate extent of pulmonary involvement and cavitary disease
    • Culture and Drug Susceptibility Testing (DST) - confirmation and detailed resistance profile
    • HIV testing - to assess TB-HIV co-infection status
    • Liver Function Tests (LFTs) - baseline assessment before anti-TB therapy initiation
    • Renal Function Tests - baseline assessment before anti-TB therapy
    • Monthly sputum AFB smear microscopy - assess treatment response (negative at 2 months expected)
    • Repeat TB-PCR at 2-3 months - assess molecular cure
    • If MTB DNA POSITIVE with Rifampicin RESISTANT (MDR-TB):
    • Extended Culture and DST - determine susceptibility to second-line drugs (fluoroquinolones, injectables)
    • Line Probe Assay (LPA) or GenoType MTBDRplus - comprehensive resistance profile
    • Baseline audiometry - before starting injectable agents
    • Baseline vision testing - before fluoroquinolone therapy
    • Electrolyte panel and renal function - for injectable drug monitoring
    • ECG - baseline before bedaquiline therapy
    • Monthly sputum AFB and TB-PCR - for treatment monitoring (longer duration required)
    • If MTB DNA NEGATIVE:
    • Repeat TB-PCR or culture if clinical suspicion remains high
    • Investigate for alternative diagnoses (bacterial pneumonia, fungal infection, malignancy)
    • Chest imaging if not already performed
    • Ongoing Monitoring Frequency:
    • Drug-Sensitive TB: Monthly sputum examination for first 3-4 months, then at end of treatment
    • MDR-TB: More frequent monitoring - sputum AFB at each visit until conversion achieved
    • Post-treatment follow-up: At 3, 6, 12 months after completion to detect relapse
  • Fasting Required?
    • NO - Fasting is NOT required
    • Specimen Type:
    • Early morning sputum (preferred) - first sputum sample after waking; has higher bacterial yield
    • May also use: Bronchoalveolar lavage, cerebrospinal fluid, pleural fluid, lymph node biopsy, other body fluids
    • Special Preparation Instructions:
    • Sputum collection: Patient should rinse mouth with water (no mouthwash) before collection
    • Cough deeply to produce sputum (minimum 2-3 ml recommended) - avoid contamination with saliva
    • Collect in sterile container provided by laboratory
    • Close container immediately and label with patient information
    • Timing of Specimen Collection:
    • Early morning sample is preferred - higher yield of TB bacilli
    • Collect on any day of the week, any time is acceptable (though early morning preferred)
    • Medications to Avoid:
    • None - no medications need to be avoided specifically for specimen collection
    • Note: If already on anti-TB treatment, specimen can still be collected and tested
    • Transport and Storage:
    • Transport to laboratory immediately or within 2-4 hours at room temperature
    • If refrigerated, store at 2-8°C; may be stored up to 7 days with refrigeration
    • Avoid freezing as it may reduce organism viability
    • Additional Patient Information:
    • Fasting is not required - patient can eat and drink normally
    • No special diet restrictions before specimen collection
    • Avoid using mouthwash or throat lozenges immediately before sputum collection
    • Smoking status should be documented - may affect sample quality

How our test process works!

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