REF 42 048 / 42 615 / 42 627 / 42 727 Half-Fraser ... - bioMerieux

REF 42 048 / 42 615 / 42 627 / 42 727 08491 F - en - 2007/09
Half-Fraser Broth (FRASER 1/2)
Selective pre-enrichment of Listeria from food and environmental specimens
SUMMARY AND EXPLANATION
Half-Fraser Broth is used for the selective pre-enrichment
of Listeria from food and environmental specimens.
The enrichment phase complies with the standard EN ISO
11 290-1 and amendment A1 (4) for the detection of
Listeria monocytogenes in foods.
PRINCIPLE
The broth contains a rich nutritive base, consisting of a
mixture of peptones, and a buffer that maintains the pH
close to neutral to favor the growth of the main species of
Listeria.
The broth is made selective by the incorporation of
lithium, actiflavin and nalidixic acid (1). Therefore, it is
particularly efficient for products with a high concentration
of microbes (3). The formulation of Half-Fraser (half
concentration in antibiotics and acriflavin) has been
developed to limit the selective capacity of the original
medium (2), in order to allow better growth of stressed
Listeria.
CONTENT OF THE KIT
Ready-to-use medium
Bottles:
REF 42 048 6 x 225 ml bottles
(250 ml bottle - FRASER 1/2 – 6F)
REF 42 615 * 4 x 225 ml bottles
(300 ml bottle - FRASER 1/2 – 4F)
* For direct inoculation
Bags / Minibags:
REF 42 627 Pack of 3 x 3 liter bags
Packaged in tinted sachets.
(FRASER 1/2 – 3P)
REF 42 727 Pack of 10 x 225 ml minibags
(FRASER 1/2 - MNB)
COMPOSITION
Theoretical formula.
This medium can be adjusted and/or supplemented according to the
performance criteria required:
Meat peptone (bovine or porcine) .................................................5 g
Casein peptone (bovine)...............................................................5 g
Meat extract (bovine or porcine) ...................................................5 g
Yeast extract.................................................................................5 g
NaCl ...........................................................................................20 g
Buffer mixture ........................................................................13.35 g
Esculin..........................................................................................1 g
Lithium chloride ............................................................................3 g
Ammonium ferric citrate.............................................................0.5 g
Acriflavin..............................................................................0.0125 g
Nalidixic acid............................................................................0.01 g
Purified water.................................................................................1 l
pH 7.3
bioMérieux ® SA English - 1
For microbiological control only
REAGENTS AND MATERIAL REQUIRED BUT NOT
PROVIDED
Reagents:
• FRASER Broth (Ref. 42 072).
• PALCAM Agar (Ref. 43 559).
• Oxford Agar (Ref. 43 560).
• chromID Ottaviani Agosti Agar (Ref. 43 641).
Material:
• Bacteriology incubator.
• Stomacher ® Bag.
POSSIBLE ADDITIONAL REAGENTS
• VIDAS ® Listeria (Ref. 30 700).
• VIDAS ® Listeria monocytogenes II (Ref. 30 704).
WARNINGS AND PRECAUTIONS
• For microbiological control only.
• For professional use only.
• This kit contains products of animal origin. Certified
knowledge of the origin and/or sanitary state of the
animals does not totally guarantee the absence of
transmissible pathogenic agents. It is therefore
recommended that these products be treated as
potentially infectious, and handled observing the usual
safety precautions (do not ingest or inhale).
• All specimens, microbial cultures and inoculated
products should be considered infectious and handled
appropriately. Aseptic technique and usual precautions
for handling the bacterial group studied should be
observed throughout this procedure. Refer to
"CLSI ® /NCCLS M29-A, Protection of Laboratory
Workers from Occupationally; Acquired Infections;
Approved Guideline – Current revision". For additional
information on handling precautions, refer to "Biosafety
in Microbiological and Biomedical Laboratories –
CDC/NIH – Latest edition", or the current regulations in
the country of use.
• Listeria monocytogenes may cause serious illness,
particularly in certain groups at risk such as pregnant
women, immunocompromised patients and the elderly.
• Culture media should not be used as manufacturing
material or components.
• Do not use broths after the expiry date.
• Do not use broths which show signs of contamination.
• Before use, make sure the tamper-proof systems are
intact (capsule, seal, stopper).
• For the 3-liter bags, only pierce the rubber stopper once.
• The medium should be used according to the procedure
indicated in this package insert. Any change or
modification in the procedure may affect the results.

Half-Fraser Broth (FRASER 1/2) 08491 F - en - 2007/09
STORAGE CONDITIONS
• Store the Half-Fraser broths at 2-8°C in their box
until the expiry date.
• After opening, the bags can be stored for 24 hours at
2-8°C.
SPECIMENS
Follow the recommendations in the current standards to
perform specimen collection and preparation.
INSTRUCTIONS FOR USE
Allow the Half-Fraser broths to come to room
temperature.
• For 3-liter bags
1. Homogenize the bag.
2. Remove the tamper-proof seal.
3. Delicately perforate the rubber stopper using a trocar
in order to pour the broth.
Identify the samples using the removable labels on the
bag.
• For MINIBAGS
MINIBAGS can be used:
- either by transferring the broth into a Stomacher ® bag
containing the test sample,
- or by directly inoculating a liquid sample.
In this case, put the stopper back in place and
incubate the MINIBAG.
Pre-enrichment:
A primary 1/10 dilution of the sample is generally
performed (e.g.: 25 g of sample in 225 ml of medium)
directly in the bottle (ref. 42 615), in the MINIBAG
(ref. 42 727) or in the Stomacher bag.
After mixing, incubate at 30°C for 24 ± 2 hours. After preenrichment,
subculture 0.1 ml in 10 ml of Fraser Broth.
Incubate at 37°C for 48 hours. After pre-enrichment and
enrichment, isolate on selective chromID Ottaviani Agosti
Agar in parallel with a medium of choice (Oxford or
PALCAM).
Notes:
• If pre-enrichment is performed in a Stomacher bag, it is
preferable to use Half-Fraser Broth 6x225 ml bottles
(ref. 42 048) or MINIBAG 10x225 ml (ref. 42 727) or
3-liter bags (ref. 42 627).
• Liquid samples can be directly inoculated in the Half-
Fraser Broth bottles (ref. 42 615) or the MINIBAGS
(ref 42 727). The volume of air in these containers
allows sufficient oxygenation of the medium.
Use with the VIDAS ® system
VIDAS is an automated immunoanalyzer for the rapid
detection of pathogenic micro-organisms in foods.
• See the recommendations in the VIDAS assay package
inserts.
• bioMérieux recommends the use of this medium to
ensure that optimum and within-run reproducible results
are obtained on the VIDAS system
bioMérieux ® SA English - 2
READING AND INTERPRETATION
Refer to the indications in the corresponding reagent
package insert.
QUALITY CONTROL
Half-Fraser Broth is designed and developed to meet the
strictest quality requirements.
The results of the strains tested in the batch by batch
quality control are given on the quality control certificate
available on request.
The quality control is performed according to the
recommendations of the standard XP CEN ISO/TS
11133-2 (5).
LIMITATIONS OF THE METHOD
• It has been found that Half-Fraser Broth is not suitable
for the enrichment of some strains of Listeria grayi. This
non-pathogenic species is mainly found in soil and is
rarely isolated from food samples.
• The broth may show a slightly greenish precipitate that
does not alter its performance.
WASTE DISPOSAL
Dispose of used and unused reagents as well as any
other contaminated disposable materials following
procedures for infectious or potentially infectious products.
It is the responsibility of each laboratory to handle waste
and effluents produced according to their nature and
degree of hazardousness and to treat and dispose of
them (or have them treated and disposed of) in
accordance with any applicable regulations.
LITERATURE REFERENCES
1. FRASER J.A., SPERBER W.H. - Rapid Detection of Listeria
spp. in Food and Environmental Samples by Esculin
Hydrolysis - J. of Food protection, 1988,vol. 51, p.762-765.
2. HOLBROOK R., ANDERSON J.M., BRIGGS T.A. et al.-
Faster detection of Listeria in food using rapid immunoassay
following culture - 3rd World Congress foodborne infections
and intoxications, (16-19 June 1992), Berlin, p.1208-1210.
3. RODRIGUEZ L, FERNANDEZ G., GARAYZABAL J. et al. -
New Methodology for the Isolation of Listeria Microorganisms
from Heavily Contaminated Environments. - Applied and
Environmental Microbiology, 1984, vol. 47, p. 1188-1190.
4. EN ISO 11290-2: 1998
Microbiology of food and animal feeding stuffs. Horizontal
method for the detection and enumeration of Listeria
monocytogenes. Part 2: Enumeration method.
EN ISO 11290-2 / A1 : 2005 Amendement 1
5. XP CEN ISO/TS 11133-2 (January 2004) – Microbiology of
food and animal feeding stuffs. Guidelines on preparation
and production of culture media. Part 2: Practical guidelines
on performance testing of culture media.

Half-Fraser Broth (FRASER 1/2) 08491 F - en - 2007/09
INDEX OF SYMBOLS
Symbol Meaning
GB : Catalogue number
US : Catalog number
Manufacturer
Temperature limitation
Use by
Batch code
Consult Instructions for Use
Contains sufficient for tests
WARRANTY
bioMérieux disclaims all warranties, express or implied,
including any implied warranties of MERCHANTABILITY
AND FITNESS FOR A PARTICULAR USE. bioMérieux
shall not be liable for any incidental or consequential
damages. IN NO EVENT SHALL BIOMERIEUX’S
LIABLITY TO CUSTOMER UNDER ANY CLAIM
EXCEED A REFUND OF THE AMOUNT PAID TO
BIOMERIEUX FOR THE PRODUCT OR SERVICE
WHICH IS THE SUBJECT OF THE CLAIM.
CLSI is a used, pending, and/or registered trademark belonging to Clinical and Laboratory Standards Institute Inc.
Stomacher is a used, pending, and/or registered trademark belonging to Seward Ltd.
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