Detection of C. botulinum (in vivo reference method)

Description of the Test

Details:

Detection and typing of Clostridium botulinum in feces

This test consists of three phases:

1. Culture of the organisms under anaerobic conditions
2. Detection and typing of genes encoding neurotoxins on DNA extracted from the cultures using real-time PCR
3. Detection and determination of the toxin type based on the mouse bioassay: identification of the toxins present in the culture supernatant by means of a lethality test and seroneutralization.

Test Results:

• Clostridium botulinum: detected / not detected
• Neurotoxin type in case of botulism: A, B, E, or F

Purpose of the Test

Detection and typing of Clostridium botulinum strains producing neurotoxins A, B, E, or F in feces

Criteria for Performing this Test within the Framework of Reference Activities

• Laboratory confirmation of clinical cases of botulism
• Typing of the neurotoxin responsible for botulism

Sample Instructions

• Type: Feces
• Minimum quantity: 15 g (2 g for infants)
• Note: In cases of suspected botulism, it is strongly recommended to provide both a serum and a fecal sample to the NRC. Serum samples should be collected as soon as possible after the onset of symptoms and before administration of antitoxins.

Transport Instructions

• Transport in appropriate packaging: https://www.biosafety.be/content/transport-annexes#6
• Transport at room temperature or in a cool box within <48 hours.

Unacceptable Requests

Damaged containers or samples that have leaked during transport.

Turnaround Time (and Analysis Frequency)

• Turnaround time: 15 working days
• Analysis days: Monday–Tuesday–Wednesday–Thursday–Friday

Reporting of Test Results

• A written report of the test results will be provided according to the analysis request via email, fax, postal mail, or eHealthBox.
• If analysis is not possible, a test report will be issued with the statement “not analyzed” and an explanatory comment.