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Utah Public Health Laboratory

Clostridium botulinum culture and toxin (Botulism)

Fee: Please refer to fee schedule

Ordering Recommendation

All clients – Contact Utah Public Health Laboratory prior to submitting specimens. Testing for any patient over the age of 1 must be approved by Utah Department of Health Epidemiology.

Performed

Sunday - Saturday as required.

Methodology

LRN Protocols

Reported

2 - 7 days

Specimen Required

  • Collect: Stool, enema fluid, gastric aspirate, vomitus, serum, tissue, wound, exudates, organism isolate, postmortem specimens, food and environmental samples.:
  • Specimen Preparation:
    • Feces: Place into sterile unbreakable container and label carefully. Confirmatory evidence of botulism may be obtained from 10-50 gram quantities (Walnut size); botulism has been confirmed in infants with only “pea-sized” stool samples. The specimen must be kept cool or refrigerated, do not freeze unless an unavoidable delay of several days is anticipated. Freezing does not affect the ability to detect toxin, but does affect the ability to detect the organism.
    • Enema: Place approximately 20 ml into sterile unbreakable container and label carefully. If an enema must be given because of constipation, a minimal amount of fluid (preferably non-bacteriostatic water) should be used to obtain the specimen so that the toxin will not be unnecessarily diluted. Transport in a Port-A-Cul vial to maintain anaerobiosis. Specimens must be kept cool or refrigerated.
    • Gastric aspirate or vomitus: Place approximately 20 ml into sterile unbreakable container and label carefully. Transport in a Port-A-Cul vial to maintain anaerobiosis. Specimens must be kept cool or refrigerated.
    • Serum: Use red top or separator type tubes to obtain serum (no anticoagulant). Samples should be obtained as soon as possible after the onset of symptoms and before antitoxin is given. Enough blood should be collected to provide at least 10 mL of serum (approximately 20 mL of whole blood). Serum volumes less than 3 ml will provide inconclusive results. Whole blood should not be sent as it typically undergoes excessive hemolysis during transit. Specimen should be kept cool or refrigerated, do not freeze unless an unavoidable delay of several days is anticipated. Note: Serum is not an acceptable sample for infant botulism testing
    • Tissue, wounds, or exudates: Place into sterile unbreakable container and label carefully. Specimens should be placed in Port-A-Cul vials and sent to the appropriate laboratory, preferably without refrigeration, for attempted isolation of C. botulinum. Swabs of superficial wounds are not acceptable for anaerobic culture. Maintain specimens at room temperature.
    • Postmortem: Obtain specimens of intestinal contents from different levels of small and large intestines. Place approximately 10 grams per specimen into sterile unbreakable container and label carefully. Obtain gastric content, serum and tissue is/as appropriate.
    • Culture: Ship suspicious isolates anaerobically (overlay liquid media with 2-inch layer of sterile petroleum jelly; melt/temper prior to overlaying culture). Cultures may be shipped at room temperature or refrigerated.
    • Food specimens: Foods should be left in their original containers if possible, or placed in sterile unbreakable containers and labeled carefully. Place containers individually in leakproof containers (i.e., sealed plastic bags) to prevent cross-contamination during shipment. Empty containers with remnants of suspected foods can be examined. Foods most likely to allow growth of C. botulinum will have a pH of 3.5-7.0 (usually 5.5-6.5). Possible foods include:
    •  Home canned products having a low acidity (pH of 4.6 or greater)
    •  Foods with low salt or low sugar content
    •  Foods that are held at temperatures that allow the organism to grow (optimal 35°C, but as low as 15°C)
    •  Foods that are consumed without prior heating.
    •  Foods that are commercially processed are rarely incriminated; however, the threat to public health is much greater with a commercial foodstuff. Unopened containers are to be sent to the U.S. Food and Drug administration (FDA), with prior arrangement. Keep the samples cool or refrigerated, do not freeze.
    • Swab samples: Send swabs in an anaerobic transport medium (e.g., Port-A-Cul tubes). For aerosolized botulinum toxin exposure, obtain nasal swabs for culture for C. botulinum. For toxin testing, serum should be used. Swabs may be shipped at room temperature or refrigerated.
  • Storage/Transport Conditions:For transport time >1h and <24h, refrigerate at 2 to 8°C. Culture isolates may be transported at room temperature. Specimens that are frozen must remain frozen until it is time to perform the test.
  • Unacceptable Conditions:Serum specimens on patients less than 1 year of age.
  • Remarks:
  • Stability:Transport directly to laboratory at room temperature. For transport time >1 h and < 24 h, refrigerate at 2 to 8°C.

Results

Detected or Not Detected

Recovered or Not Recovered

Interpretive Data

Clostridium botulinum toxin detected or not detected, toxin type

Clostridium botulinum recovered or not recovered from culture.

Note

It is mandatory that UPHL be contacted prior to submitting samples for testing

Cross References

American Society of Microbiology Sentinel Level Clinical Laboratory Guidelines http://www.asm.org/index.php/guidelines/sentinel-guidelines