VIDAS Salmonella (SLM) Assay - AOAC 996.
08
SCOPE
This method is applicable to:
− Raw meats, meat products and carcass swabs.
PRINCIPLES
Detection of Salmonella is based on enzyme-linked fluorescent immunoassay performed in
an automated VIDAS instrument. Following enrichment (see below) Salmonella is detected
in boiled broth by specific monoclonal antibodies. A fluorescent marker is then added and
Salmonella detected automatically by the VIDS machine. The detection protocol can be
broken down as follows:
▪ Pre-enrichment in non-selective liquid medium
Sample is diluted 1:10 in pre-warmed (room temperature or 37 ± 1C for large
volumes) buffered peptone water and pre-enriched at 37 ± 1C for 18 h ± 2 h. For
carcass sponges, buffered peptone water is added to the moistened sponge to bring the
total volume to 60-100 mL and the sample incubated at 37 ± 1C for 18 ± 2 h. In the case
of sponges BPW need not be warmed to room temperature before being used to re-
hydrate the sponge, for all subsequent additions BPW should be warmed to room
temperature.
▪ Selective Enrichment
Selenite cystine broth and tetrathionate broth are inoculated with pre-enrichment broth
and incubated at 35 ± 1 and 42 ± 1C, respectively, for 18 ± 2 h.
▪ Post-enrichment
Selective enrichment broths are inoculated into M-broth and incubated for 6 h at 42 ±
1C.
▪ Enzyme immunoassay
Salmonella is assayed by testing a portion of boiled post-enrichment broth. The assay is
automated and carried out in the VIDAS instrument following the manufacturer’s
instructions. The assay should be finished in approximately 45 minutes.
▪ Cultural confirmation
All Salmonella positive samples should be confirmed using refrigerated M broth
following AS 5013.10.
Issue 2023 05 26 | Approved Methods Manual
Export Standards Branch | Exports and Veterinary Services Division Page 1 of 2
Department of Agriculture, Fisheries and Forestry
Salmonella Detection-VIDAS SLM – AOAC 996.08
CHECKLIST
Pre-enrichment Is the buffered peptone water used for pre-enrichment?
Is BPW warmed prior to use (room temperature or 37 ±
1C)?
Is the correct amount of enrichment broth used for the
weight of sample analysed?
Is a positive control run with each batch of samples
analysed?
Are reference cultures inoculated into primary
enrichment broth at a level of 10 to 100 cells?
Is pre-enrichment done at 37 ± 1C for 18 h?
Selective- Is selective enrichment carried out in SC and TT broth?
enrichment
Is SC broth incubated at 35 ± 1C for 18 h?
Is TT broth incubated at 42 ± 1C for 18 h
Post-enrichment Is M-broth incubated at 42 ± 1C for 6 h?
Enzyme Are the manufacturer’s instructions available?
immunoassay
Are kits stored at 2-8C when not in use?
Cultural Is Salmonella confirmed from M-Broth or selective
confirmation enrichment media?
If an external laboratory is used is it department
(if applicable)
approved?
M broth should be supplied to off-site laboratories for
confirmation following AS 5013.10
Are Salmonella confirmed using AS 5013.10 (with
appropriate selective agar plates)?
Issue 2023 05 26 | Approved Methods Manual
Export Standards Branch | Exports and Veterinary Services Division Page 2 of 2
Department of Agriculture, Fisheries and Forestry
