Botulinum Neurotoxins

Molecular Control
Better Food. Better Health. Better World. Service DataSheet
Overview
Service Description Molecular Detection of botulinum neurotoxins

Test Code BIOMOL-BONTXNN

Testing Lab MERIEUX NUTRISCIENCES – ARABA, VITORIABIDEA 14 – 01010 VITORIA-GASTEIZ (SPAIN)

Botulinum neurotoxin, known also as Botox, is a potent neurotoxin produced by the bacterium
Clostridium botulinum. While primarily recognized for its use in cosmetic procedures to reduce
wrinkles, in the food industry, botulinum neurotoxin is of utmost concern due to its ability to cause
botulism, a severe and potentially fatal illness.

Botulism is primarily associated with improperly preserved or processed foods, especially those
with low acidity and anaerobic conditions that favor the growth of Clostridium botulinum and
subsequent toxin production.

The sensitivity, specificity, and reliability of the analytical methods for Botulinum neurotoxin are
essential for effective monitoring and control. Laboratories and food manufacturers must adhere to
stringent quality assurance protocols and validation procedures to ensure the accuracy and
precision of toxin detection. Regular surveillance and testing programs result a key step identify
potential sources of contamination and ensure the safety of food products.

The quantitative polymerase chain reaction (qPCR) approach has demonstrated several notable
improvements over other analytical options for the detection and quantification of botulinum
neurotoxin (BoNT) in various samples. Here are some key advantages of the qPCR approach,
between can be highlighted the following:

1. Sensitivity and specificity: qPCR offers high sensitivity and specificity in detecting BoNT.
Background ensuring accurate and reliable identification of BoNT presence in samples.
2. Rapid detection: qPCR enables rapid detection of BoNT compared to traditional methods.
3. Quantitative analysis: As the name suggests, qPCR allows for quantitative analysis of
BoNT. By comparing the amplification signal to a standard curve, the technique can
determine the initial amount of target DNA in the sample, providing a quantitative
measure of BoNT levels. This information is crucial for assessing the severity of
contamination and making informed decisions regarding food safety.
4. Specific serotype identification: Some qPCR assays are designed to distinguish between
different serotypes of BoNT (e.g., A, B, E and F) present in the sample. This capability is
valuable as different serotypes have varying toxicities and clinical implications. Accurate
serotype identification aids in appropriate treatment and management of botulism cases.
5. Early detection of toxin production: qPCR can detect the presence of BoNT genes even
before toxin production occurs. This early warning capability allows for proactive
measures to be taken to prevent toxin formation and reduce the risk of botulism
outbreaks.
6. Improved sample handling: Compared to traditional culture-based methods, qPCR
requires minimal sample handling and reduces the risk of laboratory-acquired infections.
This advantage not only enhances safety but also streamlines the testing process, making
it more efficient and cost-effective.
7. Compatibility with complex matrices: qPCR is compatible with various sample matrices,
including food samples, environmental samples, and clinical specimens. This versatility
allows for comprehensive monitoring and surveillance of BoNT in different settings.

Our internal method, validated according to ISO 16140:4 is optimized for the following matrices:
1. Any type of canned and preserved foods: Improperly processed or home-canned low-acid
Matrices
foods, such as vegetables, meats, fish, and soups, can create an anaerobic environment
favorable for the growth of Clostridium botulinum and toxin production.
2. Any type of Vacuum-sealed and sous vide products: Vacuum packaging and sous vide

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 Botulinum Neurotoxins
Molecular Control
Better Food. Better Health. Better World. Service DataSheet
cooking methods can create anaerobic conditions that support the growth of C.
botulinum.
3. Fermented and cured foods: Some traditional fermented and cured foods, such as
fermented fish, sausages, and raw salted meats, can be potential sources of BoNT
contamination if not prepared and stored correctly. Fermentation and curing processes
may not always eliminate the toxin if present.
4. Infused oils and flavored oils: Homemade infused oils, especially those containing garlic
or herbs, can pose a risk of botulism if not prepared and stored properly. The low-acid,
anaerobic environment in the oil can support C. botulinum growth and toxin production.
5. Honey and other sweeteners: Honey has been associated with infant botulism, a form of
botulism that affects young infants. Honey may contain spores of C. botulinum, which can
multiply and produce toxins in the infant's immature digestive system. Other sweeteners,
such as corn syrup, can also be potential sources of contamination if improperly
processed.

Always check matrices and prices with your Customer Care (CC) representative.
Due to complexity and variability a sample might require a:
(*) Special Notes  Short validation
 Revision of the limits
to be always confirmed by the corresponding CC

The molecular methodology for this assay focuses on targeting a specific genomic region associated
with botulinum neurotoxin (BoNT) production. The assay employs primers and probes designed to
amplify and detect conserved regions within the genetic material of the BoNT gene.
Method Description
The target genomic region depends on the specific serotype(s) of BoNT being investigated.
Different serotypes, such as A, B, E, etc., have unique genetic sequences that distinguish them from
one another. Therefore, the assay design considers these serotype-specific variations to ensure
accurate detection and discrimination.

Technical Data
Instrument PCR Real Time thermocycler without requirement of passive reference

Limit of Quantification/ While there is no universally applicable LOD for qPCR-based assays of all BoNT serotypes, the
Detection general LODs for this assay is in the range of 10 to 100 copies of the target gene per reaction.

Turn Around Time (TAT)

Standard detection: 5-7 working days.

Standard TAT
RUSH Service: 3 working days only after agreement with laboratory.
(rush TAT if present)
For quantification always confirm delivery time with laboratory.

Sampling
Sample Quantity At least 50 gr.

Sampling Procedure For fresh sample: storage and transportation at 4ºC

In case of non-canned/ packed samples extreme caution to avoid possible cross-contamination of

(*) Special Notes the samples.
In case of doubt consult with your Customer Care (CC) representative.

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