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1

The 3M Molecular Detection Assay 2 –

Listeriamonocytogenes

method may generate

Listeria

2

monocytogenes

to levels sufficient to cause stillbirths and fatalities in pregnant women and the

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immunocompromised, if exposed.

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To reduce the risks associated with exposure to chemicals and biohazards:

It is strongly recommended that female laboratory staff be informed of the risk to a

developing fetus resulting from infection of the mother through exposure to

Listeria

monocytogenes

Perform pathogentesting in a properly equipped laboratory under the control of trained

personnel

Always follow standard laboratory safety practices, including wearing appropriate protective

apparel and eye protection while handling reagents and contaminated samples

Avoid contact with the contents of the enrichment media and reagent tubes after amplification

Dispose of enriched samples according to current industry standards

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Enrichment

Sample Preparation

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3M recommends the use of Demi-Fraser Broth (with addition of ferric ammonium citrate) for the

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enrichment of food and environmental samples.Table 1 presents guidance for the enrichment of

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food and environmental samples. It is the user’s responsibility to validate alternate sampling

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protocols or dilution ratios to ensure this test method meets the user’s criteria.

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Foods

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a)

Allow the Demi-Fraser Broth enrichment medium to equilibrate to ambient laboratory

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temperature.

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b)

Aseptically combine the enrichment medium and sample according to Table 1. For all meat

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and highly particulate samples, the use of filter bags is recommended.

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c)

Homogenize thoroughly by blending, stomaching, or hand mixing for 2 ±0.2 minutes.

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Incubate at 37 ±1°C according to Table 1.

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Environmental samples

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Sample collection devices can be a sponge

hydrated

with a neutralizing solution to inactivate the

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effects of the sanitizers. 3M recommends the use of a biocide-free cellulose sponge. Neutralizing

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solution can be Dey-Engley (D/E) Neutralizing Broth or Letheen broth. It is recommended to

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sanitize the area after sampling.

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The recommended size of the sampling area

for verifying

the presence or absence of the pathogen

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on the surface is at least 100 cm

2

(10 cm x 10 cm or 4”x4”). When sampling with a sponge,

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cover the entire area going in two directions (left to right then up and down) or collect

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environmental samples following your current sampling protocol or according to the FDA BAM,

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USDA FSIS MLG or ISO 18593 (7) guidelines.

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1.

Allow the Demi-Fraser Broth enrichment medium to equilibrate to ambient laboratory

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temperature.

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2.

Aseptically combine the enrichment medium and sample according to Table 1.

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3.

Homogenize thoroughly by blending, stomaching, or hand mixing for 2 ±0.2 minutes.

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AOAC Research Institute

Expert Review Panel Use Only

OMAMAN-30 D/ PTM Validation Report 081501

OMA ERP - June 2016

ERP Use Only