Guidelines for Specimen Collection and Submission: Mycobacteriology (AFB) Collection, Transport, and Storage

Quick Links

Specimen Type (Reference): Abscess - General

Collection guidelines: Remove surface exudate by wiping with sterile saline or 70% alcohol. Collect fluid abscess material with Luer tip syringe and/or remove tissue aseptically.

Transport device and/or minimum vol.:   Fluid abscess material should be submitted in a sterile leak-proof container such as a sterile 50 ml conical tube.  Tissue should be submitted in 2 to 3 ml sterile saline in a sterile leak-proof container such as a sterile 50 ml conical tube. Do not transport specimen in culturette or in transport medium. If transport is delayed for more than one hour, refrigerate specimen

Transport time and temp:  As soon as possible; Overnight shipment on cold pack is recommended. 

Storage time:  Refrigerate, but do not freeze specimen

Replica limits:  1/day/source

Comments: Tissue (at least 1 gram, if possible) or fluid is preferred.  A swab is discouraged unless it is the only specimen available; submit swabs in 2 to 3 ml sterile saline.  Swabs submitted in transport medium or culturette tube are unacceptable.  Do not freeze or preserve tissue.

^  Top

Specimen Type (Reference): Abscess - Open

Collection guidelines: Aspirate if possible.  Aspirate material from under the margin of the lesion/abscess.  Remove tissue aseptically.

Transport device and/or minimum vol.:  Fluid abscess material should be submitted in a sterile leak-proof container such as a sterile 50 ml conical tube.  Tissue should be submitted in 2 to 3 ml sterile saline in a sterile leak-proof container such as a sterile 50 ml conical tube. Do not transport specimen in culturette or in transport medium.

Transport time and temp:  As soon as possible; Overnight shipment on cold pack is recommended. If transport is delayed for more than one hour, refrigerate specimen.

Storage time: Refrigerate but do not freeze specimen

Replica limits: 1/day/source

Comments: Tissue (at least 1 gram, if possible) or fluid is preferred.  A swab is discouraged unless it is the only specimen available; submit swabs in 2 to 3 ml sterile saline.  Swabs submitted in transport medium or culturette tube are unacceptable.  Do not freeze or preserve tissue.

^  Top

Specimen Type (Reference): Body Fluids: abdominal, amniotic, ascites, bile, joint, paracentesis, pericardial, peritoneal, pleural, synovial, thoracentesis 

Collection guidelines: Palpate vein before disinfection of venipucture site. Disinfection of venipuncture site: 

  1. Disinfect overlying skin with alcohol or iodine preparation  
  2. Obtain specimen via percutaneous needle aspiration or surgery 
  3. Always submit as much fluid as possible; never submit a swab dipped in fluid 

Transport device and/or minimum vol:  Sterile leak-proof container such as a 50 ml conical tube; Minimum volume is 10-15 ml.

Transport time and temperature: As soon as possible; Overnight shipment on cold pack is recommended. If transport is delayed for more than one hour, refrigerate specimen.  

Storage time:  Refrigerate but do not freeze specimen

Replica limits: None

Comments: Maximum volume recommended is 15 ml

^  Top

Specimen Type (Reference): Cellulitis, aspirate from area of

Collection guidelines:

  1. Cleanse site by wiping with sterile saline or 70% alcohol.
  2. Aspirate the area of maximum inflammation (commonly the center rather than the leading edge) 
  3. with a needle and syringe; irrigation with a small amount of sterile saline may be necessary
  4. Aspirate saline into syringe, and expel into sterile leak-proof container such as a sterile 50 ml conical tube

Transport device and/or minimum vol.: Sterile leak-proof container such as a sterile 50 ml conical tube

Transport time and temperature:  As soon as possible; Overnight shipment on cold pack is recommended. If transport is delayed for more than one hour, refrigerate specimen.

Storage time: Refrigerate, but do not freeze specimen

Replica limits: 1/day

Transport time and temp:  As soon as possible; Overnight shipment on cold pack is recommended. If transport is delayed for more than one hour, refrigerate specimen.

^  Top

Specimen Type (Reference): Cerebrospinal Fluid (CSF) Analysis

Collection guidelines: 1.  Disinfect site with iodine preparation

Transport device and/or minimum volume: Sterile leak-proof container such as 50 ml conical tube.  Minimum amt required:  2-3 ml

Transport time and temp: As soon as possible; Ambient

Storage time: Room Temperature.  Do not refrigerate or freeze CSF specimen.

Replica limits: None

Comments: None

^  Top

Specimen Type (Reference): Endometrial Tissue

Collection guidelines: Collect tissue aseptically during endometrial biopsy procedure. 

Transport device and/or minimum vol: Sterile leak-proof container; if dry, add up to 5 ml sterile saline for transport.

Transport time and temp: Overnight shipment on cold pack is recommended. If transport is delayed for more than one hour, refrigerate specimen.

Storage time: Refrigerate, but do not freeze specimen.

Replica limits: 1/day 

Comments: Do not freeze or preserve tissue specimen.

^  Top

Specimen Type (Reference): Eye - Corneal Scrapings 

Collection guidelines:

  1. Specimen collected by ophthalmologist
  2. Using sterile spatula, scrape ulcers or lesions, and inoculate scraping directly onto medium 

Transport device and/or minimum vol.: Direct culture inoculations:  BHI with 10% sheep blood agar plate or chocolate agar plates 

Transport time and temp: As soon as possible; Overnight shipment on cold pack is recommended. If transport is delayed for more than one hour, refrigerate specimen.

Storage time: Refrigerate but do not freeze specimen

Replica limits: None

Comments: None

^  Top

Specimen Type (Reference): Eye (exudates) 

Collection guidelines: Aspirate if possible. Collect material in Luer tip syringe.

Transport device and/or minimum volume: Exudate should be submitted in a sterile leak-proof container such as a sterile 50 ml conical tube.

Transport time and temperature: As soon as possible; Overnight shipment on cold pack is recommended.. If transport is delayed for more than one hour, refrigerate specimen 

Storage time: Refrigerate but do not freeze specimen  

Replica limits: None 

Comments: None

^  Top

Specimen Type (Reference): Feces

Collection guidelines: Pass specimen directly into a sterile leak-proof container. Do not use holding medium or transport medium. 

Transport device and/or minimum volume: Sterile, leak-proof, container such as a 50 ml conical tube; minimum volume is 1 gram. 

Transport time and temp:  Unpreserved; As soon as possible; Overnight shipment on cold pack is recommended. If transport is delayed for more than one hour, refrigerate specimen

Storage time: Refrigerate but do not freeze specimen

Replica limits: 1/Day 

Comments: Do not submit rectal swabs for mycobacterial culture. 

^  Top

Specimen Type (Reference): Gastric Wash or lavage for mycobacteria

Collection guidelines: Collect in early morning before patients eat and while they are still in bed.

  1. Introduce a nasogastric tube into the stomach
  2. Perform lavage with 25-50 ml of chilled, sterile distilled water  
  3. Recover sample and place in a leak-proof, sterile container

Transport device and/or minimum vol.: Sterile, leak-proof container such as sterile 50 ml conical tube ; minimum volume is 5-10 mL.

Transport time and temp: Neutralize within 1 h of collection, transport as soon as possible; Ambient

Storage time: NA

Replica limits: 1/day on three consecutive days

Comments: The specimen must be processed promptly, since mycobacteria die rapidly in gastric washings. Neutralize with 100 mg sodium carbonate.   Maximum volume of specimen recommended is 15 ml.

^  Top

Specimen Type (Reference): Respiratory, lower bronchoalveolar lavage, brush or wash, endotracheal aspirate, transtracheal aspirate

Collection guidelines: 

  1. Collect washing or aspirate in a sputum trap
  2. Place brush in sterile leak-proof container with up to 5 ml of saline 

Transport device and/or minimum volume: Sterile container such as a sterile 50 ml conical tube; minimum volume is 3 ml 

Transport time and temperature: As soon as possible; Overnight shipment on cold pack is recommended. If transport is delayed for more than one hour, refrigerate specimen

Storage time: Refrigerate but do not freeze specimen

Replica limits: 1/day

Comments: None

^  Top

Specimen Type (Reference): Skin Lesion

Collection guidelines:

  1. Cleanse with sterile saline, and remove lesion's surface with a sterile scalpel blade
  2. Allow transudate to accumulate 
  3. While pressing the base of the lesion, collect transudate with sterile syringe. Place contents of syringe in sterile leak-proof container.  
  4. If tissue is collected, place tissue in sterile leak-proof container and add 2 to 3 ml sterile saline for transport.

Transport device and/or minimum volume:  Sterile leak-proof container such as a sterile 50 ml conical tube. 

Transport time and temperature: As soon as possible; Overnight shipment on cold pack is recommended. If transport is delayed for more than one hour, refrigerate specimen. 

Storage time: Refrigerate but do not freeze specimen

Replica limits: 1/day

Comments:  A swab is discouraged unless it is the only specimen available; submit swabs in 2 to 3 ml sterile saline.  Swabs submitted in transport medium or culturette tube are unacceptable

^  Top

Specimen Type (Reference): Sputum, Expectorated

Collection guidelines:

  1. Collect specimen under the direct supervision of a nurse or physician 
  2. Have patient rinse or gargle with water to remove excess oral flora. 
  3. Instruct patient to cough deeply to produce a lower respiratory specimen (not postnasal fluid)
  4. Collect in a sterile container

Transport device and/or minimum volume: Sterile container such as 50 ml conical tube; minimum volume is 3 ml

Transport time and temperature: As soon as possible; Overnight shipment on cold pack is recommended. If transport is delayed for more than one hour, refrigerate specimen

Storage time: Refrigerate but do not freeze specimen

Replica limits: For diagnosis, at least three consecutive sputum specimens are needed, each collected in 8- to 24-hour intervals, with at least one being an early morning specimen.

Comments: Early morning sputum specimen is preferred. Do not pool sputum specimens.

^  Top

Specimen Type (Reference): Sputum, Induced 

Collection guidelines:

  1. Have patient rinse mouth with water after brushing gums and tongue
  2. With the aid of a nebulizer, have patients inhale approximately 25 ml of 3-10% sterile saline 
  3. Collect in a sterile leak-proof container

Transport device and/or minimum vol.: Sterile leak-proof container such as a 50 ml conical tube; minimum volume is 3 ml

Transport time and temp: As soon as possible;  Overnight shipment on cold pack is recommended.. If transport is delayed for more than one hour, refrigerate specimen

Storage time: Refrigerate but do not freeze specimen

Replica limits: For diagnosis, at least three consecutive sputum specimens are needed, each collected in 8- to 24-hour intervals, with at least one being an early morning specimen.

Comments: Same as above for sputum, expectorated.

^  Top

Specimen Type (Reference): Tissue / Lymph Node

Collection guidelines: Aseptically collected during surgery or cutaneous biopsy procedure

Transport device and/or minimum vol.: Sterile leak-proof container such as sterile 50 ml conical tube

Transport time and temp: As soon as possible; Overnight shipment on cold pack is recommended. If transport is delayed for more than one hour, refrigerate specimen.

Storage time: NA

Replica limits: 1/day

Comments: Always submit as much tissue as possible. Add 2 to 3 ml sterile saline to tissue for transport.

^  Top

Specimen Type (Reference): Vaginal Secretions

Collection guidelines:

  1. Wipe away old secretions and discharge 
  2. Obtain secretions from the mucosal membrane of the vaginal wall with a sterile pipette  

Transport device and/or minimum volume: Submit secretions in sterile leak-proof container; do not use transport medium.

Transport time and temperature: As soon as possible; Overnight shipment on cold pack is recommended.. If transport is delayed for more than one hour, refrigerate specimen

Storage time: Refrigerate but do not freeze specimen

Replica limits: 1/day

Comments: A swab is discouraged unless it is the only specimen available; submit swabs in 2 to 3 ml sterile saline. Swabs submitted in transport medium or culturette tube are unacceptable.

 

Laboratory Services

Laboratory Services

Mycobacteriology and Mycology