Approved methods for microbiological testing of meat and meat products

Amended April 2025

The following is a list of Department of Agriculture, Fisheries and Forestry (DAFF) approved test methods for meat and meat products. All testing of product relating to export certification, including carcass testing under National Carcase Microbiology Monitoring Program (formerly known as ESAM), must be by one of the methods listed with the modifications and options specified; no other modifications are permitted. Laboratory manuals and protocols must reflect the above and will be subject to audits to ensure compliance.

• Aerobic Plate Count/Total Viable Count (TVC)
• Escherichia coli O157:H7
• Shiga-toxin producing Escherichia coli  (STEC)
• Generic Escherichia coli
• Listeria
• Salmonella

Aerobic Plate Count/Total Viable Count (TVC)

• Australian Standard AS 5013.5-2016 Horizontal method for the enumeration of microorganisms – Colony count at 30°C by the pour plate technique
• AOAC 990.12 TVC Petrifilm™
• AOAC 2015.13 Neogen Petrifilm® Rapid Aerobic Count (RAC) Plate Method
• AOAC 2008.10 TEMPO TVC Method: Automated Enumeration of Total Viable Count in Food
• AOAC 010404 Compact Dry TC
• AOAC 091702 and MicroVal 2015LR52 MC-Media PAD AOAC 091702 is a validation study for incubation of MC-Media Pad at 35 ± 1°C for 24 -48 h and applies only to 50 g raw meats and other foods. MicroVal 2015LR52 is a validation study for 10 g samples, incubated at 30 ± 1°C for 72 h

Escherichia coli O157:H7

• AS 5013.26:2020 Microbiology of food and animal feeding stuffs ¡V Horizontal method for the detection of Escherichia coli O157. 
• FSIS MLG 5 Detection, isolation, and identification of Escherichia coli O157:H7 from meat products.
• FDA BAM Chapter 4A(K) Diarrheagenic Escherichia coli - Enrichment and isolation of E. coli Serotype O157:H7 from Foods

Screening methods

Where positive confirmation is required such confirmation must be by a DAFF approved confirmatory method.

• FSIS MLG 5A
  FSIS procedure for the use of Escherichia coli O157:H7 screening tests
• AOAC 031002 DuPont Qualicon BAX® System PCR Assay for Real-Time E. coli O157:H7 
• AOAC 2005.04 Assurance GDS for Escherichia coli O157:H7 in Selected Foods
• AOAC 071001 MicroSEQ(R) Real-Time PCR System for Detection of E. coli O157:H7 in raw ground beef and beef trim
• AOAC 2017.01 3M Molecular Detection Assay (MDA) 2 – E. coli O157 (including H7)
• AOAC 022002 BACGene E. coli O157:H7 Workflow
• AOAC 2019.03 GENE-UP E. coli O157:H7 2 (ECO 2)
• AOAC 021501 SureTect E. coli O157:H7 PCR Assay

The following rapid methods are not to be used for the routine testing of export meat and meat products for E. coli O157. They are approved as backup methods for use when PCR methods are temporarily unavailable. They may be used for the testing of product under commercial arrangements when a methodology is not specified under that arrangement.

• AOAC 996.09 BioControl VIP (8-12 h and 18-28 h options)
• AOAC 2000.13 Reveal (8-hours)
• AOAC 2000.14 Reveal (20-hours)
• AOAC 070201 (Performance Tested Method) Rapid√ for Escherichia coli O157 Lateral Flow Assay

Shiga-toxin producing Escherichia coli (STEC)

• MLG 5C Detection, Isolation and Identification of Top Seven Shiga Toxin-Producing Escherichia coli (STEC) from Meat and Meat Products
• FSIS MLG 5B Detection and isolation of non-O157 Shiga-toxin Producing Escherichia coli (STEC) from meat products
• AOAC 2020.06  GENE-UP EHEC Detection Method
• AOAC 081901 - NeoSeek STEC – Detection and identification of top 7 STEC

Screening methods

Where positive confirmation is required such confirmation must be by a DAFF approved confirmatory method

• AOAC 071301 Assurance GDS® MPX Top 7 STEC for detection of top 7 pathogenic STEC in beef trim. 
• AOAC 091301 DuPont Qualicon BAX® System Real-Time PCR Assays for detection of selected STEC in beef trim. 
• AOAC 0100701 IEH E. coli Test System for detection of non-O157 Shiga-toxin producing E. coli and E. coli O157 in raw ground beef. 
• AOAC 061602  RapidFinderTM STEC Detection Workflow for detection of top 7 STEC serogroups in beef products. 
• AOAC 031401 GeneDisc(R) Plate  STEC Top 7 methods for detection of O157 and top 6 non-O157 Shiga toxin producing E. coli in raw ground beef and beef trim
• AOAC 101502 Assurance GDS® MPX ID for Top 6 STEC 
• AOAC 022003 BACGene STEC Top 7 Workflow
• AOAC 071902 Neogen Molecular Detection Assay (MDA) 2 – STEC Gene Screen (stx and eae)
• MLG 5C Appendix 3, 4 and 5 - Detection, Isolation and Identification of top seven STEC from Meat & Meat Products (screening only)
• AOAC 012102 – SureTect E. coli O157:H7 and STEC Screening PCR Assay

Generic Escherichia coli

• Australian Standard AS 5013.15 General guidance for enumeration of presumptive Escherichia coli - Most probable number technique
• AOAC 991.14 E. coli PetriFilm™ Butterfields or buffered peptone water or 0.1% Peptone Salt Solution must be used as diluent
• AOAC 998.08 E. coli PetriFilm™ 
• AOAC 2005.03 SimPlate® Colour Indicator: Detection and Quantitation of Coliforms and E. coli in foods
• AOAC 2009.02 Tempo® EC AFNOR Bio 12/13 ¡V 02/05 for testing of generic E. Coli
• AOAC 110402 Compact Dry EC
• AOAC 070901 and MicroVal 2017LR71 MC-Media Pad EC . AOAC 070901 is a validation study that applies only to 50 g raw meats and other foods. MicroVal 2017LR71 is a validation study that applies to sample diluted 1:10.
• AOAC 2018.13 - Neogen Petrifilm Rapid E. coli/Coliform Count Plate

Listeria

Confirmation methods

• Australian Standard AS 5013.24.1 Food and animal feeding stuffs ¡V Horizontal method for the detection and enumeration of Listeria monocytogenes. Detection method
• FSIS MLG 8 Isolation and identification of Listeria monocytogenes from red meat, poultry, egg, and environmental samples

Note alternative secondary enrichment has been included

Screening methods

Where positive confirmation is required such confirmation must be by Australian Standard AS 5013.24.1 or FSIS MLG 8
Note the following bio-chemical test systems can be used for confirmation for all methods MICRO-ID® Listeria or API®-Listeria or VITE® 2 Compact, . β-lysin. CAMP factor discs (Remel #21-120, or equivalent) can be used instead of the traditional CAMP test procedure

• FSIS MLG 8A FSIS procedure for the use of Listeria monocytogenes BAX screening test Note the method has been updated to include testing of liquid egg products
• FDA BAM Cha 10 Detection and Enumeration of Listeria monocytogenes in Foods
• AOAC 2003.12 Automated BAX System for Detection of Listeria monocytogenes in Foods
• AOAC 999.06 VIDAS LIS Assay for Listeria in Foods (AOAC 2004.06 - VIDAS LIS modification)
• AOAC 996.14 BioControl Assurance Listeria Immunoassay
• AOAC 997.03 BioControl Listeria Visual Immunoprepicipate (VIP) Assay
• AOAC 070401 Foodproof Listeria mon9ocytogenes Detection Kit, 5¡¦Nuclease and Hybridization Probes
• AOAC 031204 Pall GeneDisc(R) method for the detection of Listeria monocytogenes in food and environmental samples
• AOAC 011022 MicroSEQ(R) Real-Time PCR System for Detection of Listeria monocytogenes in food
• NF BIO 12/33 – 05/12 VIDAS UP Listeria method (VIDAS LPT)
• Solus Listeria ELISA – NF SOL 37/02 – 06/13
• AOAC 071304 Thermo Scientific SureTect Listeria species PCR Assay (AFNOR UNI 03/09 - 11/13)
• AOAC 121402 DuPontTM BAX® System Real-Time PCR Assay for Listeria monocytogenes
• AOAC 2016.08 Neogen Molecular Detection Assay (MDA) 2 – Listeria monocytogenes Method
• AOAC 061703 BACGene Listeria monocytogenes

 Salmonella

Confirmation methods

• Australian Standard AS 5013.10 Microbiology of food and animal feeding stuffs - Horizontal method for the detection of Salmonella spp The following options are required: Second agar choice must be capable of detecting H2S negative Salmonella (internationally validated Salmonella media e.g. BGA, BGS, Rambach, ChromAgar)
• FSIS MLG 4 Isolation and identification of Salmonella from meat, poultry and egg products

Screening methods

When positive confirmation is required such confirmation must be by Australian Standard AS 5013.10 or FSIS MLG 4

• FSIS MLG 4C FSIS procedure for the use of the BAX system PCR assay for screening Salmonella in raw meat, carcass sponge samples, whole bird rinses, ready-to-eat meat and poultry products and pasteurised egg products
• AOAC 2009.03 Assurance GDS™ Salmonella method for foods
• AOAC 992.11 BioControl Assurance EIA
• AOAC 999.08 BioControl Assurance Gold pre-enrichment with BPW + novobiocin as per instructions
• AOAC 999.09 BioControl VIP
• AOAC 996.08 VIDAS Salmonella (SLM) Assay
• AFNOR BIO 12/16 ¡V 09/05 VIDAS EASY Salmonella method
• 2013.01(071101) VIDAS UP Salmonella method (VIDAS SPT)
• AOAC 2001.09 VIDAS Immuno Concentration Salmonella (ICS)
• AOAC 2009.03 Assurance GDS™ Salmonella method for foods and Assurance GDS Salmonella Tq method
• AOAC 100701 IEH PCR assay for detection of Salmonella in carcass and environmental sponges or swabs
• AOAC 120301 foodproof Salmonella Detection Kit, 5¡¦Nuclease and Hybridization Probes
• AOAC 100201 DuPont Qualicon BAX (R) System Salmonella2 PcR Assay
• AOAC 2013.02 DuPont Qualicon BAX(R) System real-time PCR assay for Salmonella
• AOAC 031001 MicroSEQ(R) Real-Time PCR System for Detection of Salmonella in food
• AOAC 050602 Assurance GDS for Salmonella
• AOAC 2014.01 Neogen Petrifilm Salmonella Express System
• AOAC 011404 Veriflow Salmonella species (SS)
• NF SOL 37/01 – 06/13 Solus Salmonella ELISA
• AOAC 2021.01 Thermo Scientific SureTect Salmonella spp PCR Assay (AFNOR UNI 03/07 – 11/13)
• AOAC 2016.01 Neogen Molecular Detection Assay (MDA) 2 – Salmonella Method​​
• AOAC 121501 BACGene Salmonella spp.