Sputum for AFB

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AFB testing may be used to detect several different types of acid-fast bacilli, but it is most commonly used to identify an active tuberculosis (TB) infection caused by the most medically important AFB, Mycobacterium tuberculosis.

Sputum: Viscous material that is derived from the lower air passages such as the lungs and bronchi that may contain substances such as mucus, blood, pus and/or bacteria; it is not the saliva that is produced by the glands in the mouth.

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SpecimenSpecimen Type: Sputum
Container/Tube: Sterile leak-proof container
Volume: 10 mL
Minimum Volume: 5 mL
MethodsZN Stain, Fluorescent Microscopy, Conventional Microscopy, Auramine Stain

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Why to Get Tested:

To help diagnose tuberculosis (TB) and infections caused by other Mycobacterium species, which are known as acid-fast bacilli (AFB), in people at risk of developing mycobacterial infections; to monitor the effectiveness of treatment

When to get Tested:

  • When you have signs and symptoms of a lung infection, such as a chronic cough, weight loss, fever, chills, and weakness, that may be due to TB or a nontuberculous mycobacterial (NTM) infection
  • When you have a positive IGRA blood test or Tuberculin skin test (TST) and you are in a high-risk group for progressing to active TB
  • When you have a skin or other body site infection that may be due to mycobacteria; when you are undergoing treatment for TB

Purpose Of The Test (Indications)

  1. This is a special stain for the diagnosis of Mycobacterium tuberculosis in various specimens like sputum, caseous material, and tissue.
  2. The AFB smear stain used to monitor the treatment of Tuberculosis.
  3. The AFB stain on sputum is indicated in a patient with:
    1. A cough.
    2. Night sweating.
    3. Anorexia.
    4. Fever.
    5. Hemoptysis.
    6. Unexplained weight loss.
    7. Fatigue and weakness.
    8. Chest pain.
  4. Advise AFB stain in high-risk patients like immunocompromised patients, alcoholics, or have recent exposure to TB patients.

Specimen Collection and Instructions

Specimen Type: Sputum
Container/Tube: Sterile leak-proof container
Volume: 10 mL
Minimum Volume: 5 mL

  1. Sputum must be first morning specimen.
  2. Prior to eating, rinse mouth with water.
  3. Expectorate into a sterile cup.
  4. Collect 5 to 10 mL of sputum in a sterile container in the morning.
  5. If the volume of sputum received is less than 1 mL, document on the report.
  6. Maximum of 3 expectorated sputum specimens, collected on separate days, will be tested on any one patient.
  7. If two specimens are received on a single day (except weekend collections), the specimens will be combined before concentration.
  8. Specimens collected by bronchoscopy will be accepted even if the patient has had 3 sputum specimens cultured prior to bronchoscopy.
  9. After therapy is initiated, subsequent cultures to follow therapy will be accepted.

Transport Temperature: Refrigerated

Test Procedure:

There are 2 Most common Procedure to Examin the Sputum smears.


Positive Result is:

  1. The acid-fast bacilli are red color rods and the background is blue.
  2. When these Rods are seen indicate active tuberculosis.
  3. Positive AFB stain may be confirmed by the culture.

The Negative Result Is:

A negative result means that:

  1. There is no infection.
  2. The number of bacteria was not in sufficient amount to be seen under the microscope.
  3. Symptoms may be due to some other cause.

Reporting Of The AFB Stain.

Presence of AF bacilliPositivity  report
When rarely seen1+ Rare
When few bacilli were seen2+ Few
When the moderate number of bacilli are seen3+  Moderate
When many bacilli are seen4+ Many
NegativeWhen no bacilli found

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