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Food Testing
Food Safety and Quality DiagnosticsBio-Rad Solutions
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Routine Testing • Quality Indicator
Contents
This booklet is only an indication provided by Bio-Rad for the use of Bio-Rad products.It is not a contractual document.
PART 1 • Routine Testing - Quality Indicator 3
Preparation of dilutions 3
Total flora 3
Escherichia coli - ISO standards 4
Escherichia coli - Alternative method: RAPID’E.coli 2 Agar 5
Coliforms - ISO standards 6
Coliforms - Alternative method: RAPID’E.coli 2 Agar 7
Enterobacteriaceae 8
Enterobacteriaceae - Alternative method: RAPID’Enterobacteriaceae 9
PART 2 • Pathogen Testing 10
Salmonella - ISO standard 10
FDA-BAM Salmonella protocol 11
FSIS Salmonella MLG 4.07 11
Salmonella - Alternative method: RAPID’Salmonella Agar double enrichment protocol 12
Salmonella - Alternative method: RAPID’Salmonella Agar short protocol 12
Salmonella - Alternative method: iQ-Check™ Salmonella II real-time PCR Kit 12
Cronobacter spp. - ISO standard 14
FDA-BAM Cronobacter spp. 14
Cronobacter spp. - Alternative method: RAPID’Sakazakii Agar short protocol 15
Cronobacter spp. - Alternative method: iQ-Check Cronobacter spp. real-time PCR Kit 15
Listeria monocytogenes - ISO standards 16
FDA-BAM Listeria monocytogenes protocol 18
Health Canada Listeria monocytogenes protocol MFHPB-30 18
Listeria monocytogenes - Alternative method: RAPID’L.mono Agar 19
Listeria monocytogenes - Alternative method: iQ-Check Listeria monocytogenes II real-time PCR Kit 20
Listeria spp. - Alternative method: RAPID’L.mono Agar 21
Listeria spp. - Alternative method: RAPID’Listeria spp. Agar 21
Listeria spp. - Alternative method: iQ-Check Listeria spp. real-time PCR Kit 22
Escherichia coli O157:H7 - Alternative method: RAPID’E.coli O157:H7 Agar 23
STEC - iQ-Check STEC VirX and SerO real-time PCR Kits 24
Escherichia coli O157:H7 - Alternative method: iQ-Check E.coli O157:H7 real-time PCR Kit 24
Campylobacter spp. - ISO standard 25
FDA-BAM Campylobacter spp. protocol 26
Campylobacter spp. - Alternative method: iQ-Check Campylobacter real-time PCR Kit 26
Campylobacter spp. - Alternative method: RAPID’Campylobacter spp. 27
Staphylococcus aureus - ISO standards 28
FDA-BAM Staphylococcus aureus 29
Staphylococcus aureus - Alternative method: RAPID’Staph Agar 29
Sulfite-reducing bacteria 30
Clostridium perfringens 30
Bacillus cereus 31
Shigella 31
Validations 32
Range for Food Testing 33
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Food Safety and Quality Diagnostics • Bio-Rad Solutions | | 3
Routine Testing • Quality Indicator
Preparation of dilutions Total flora
NF EN ISO 6887-1 (09/1999)
Preparation of test samples, initial suspension, and decimal dilutions
for microbiological examination
Part 1: General rules for preparation of dilutions for
microbiological examination
NF EN ISO 4833 (05/2003)
General guidance for the enumeration of microorganisms
Colony-count technique at 30°C
Dilution 10-1
(and following dilutions)
m g or v ml of sample+
9 x m g or 9 x v ml ofTryptone Salt or Buffered
Peptone Water (and following dilutions)
Enumeration
PCA
Initial suspensionand/or
decimal dilutions
Ordering Information
Catalog # Description
Buffered Peptone Water/BrothReady-to-use355-4179 225 ml x 6 bottles355-5795 3 L x 4 bags355-5790 5 L x 2 bags
Dehydrated356-4684 500 g
PCA (Plate Count Agar)/AgarReady-to-use355-4459 100 ml x 6 bottles355-4457 200 ml x 6 bottles356-3989 90 mm x 20 dishes
Dehydrated356-4475 500 g
Tryptone-Salt/DiluentReady-to-use355-5754 9 ml x 25 tubes355-5756 90 ml x 6 bottles355-5796 3 L x 4 bags
Dehydrated356-4544 500 g
30°C72 hrs
PCA Bacteria
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Routine Testing • Quality Indicator
4 | | Food Safety and Quality Diagnostics • Bio-Rad Solutions
ISO 16649-1 (04/2001)
Horizontal method for the enumeration of β-glucuronidase-positive E. coli
Part 1: Colony-count technique at 44°C using membranes and5-bromo-4-chloro-3-indolyl
-D-glucuronide
TBX
Direct enumeration
Sample/initial suspension
MMGA with membrane
ISO 16649-2 (04/2001)
Horizontal method for the enumerationof β-glucuronidase-positive E. coli
Part 2: Colony-count technique at44°C using 5-bromo-4-chloro-
3-indolyl -D-glucuronide
Dilutions
Resuscitation
Direct enumeration
Sample/initial suspension
Dilutions
TBX
Ordering Information
Catalog # Description
Buffered Peptone Water/BrothReady-to-use355-4179 225 ml x 6 bottles355-5795 3 L x 4 bags355-5790 5 L x 2 bags
Dehydrated356-4684 500 g
EC Medium/BrothDehydrated356-4764 500 g
Kovacs Reagent (for Indole detection)355-5313 15 ml x 2 bottles
Peptone Water (Indole Free)/BrothReady-to-use355-4180 9 ml x 25 tubes
Dehydrated356-4334 500 g
RAPID’E.coli 2/AgarReady-to-use355-5297 200 ml x 6 bottles355-5299 100 ml x 6 bottles
Dehydrated356-4024 500 g
TBX/AgarReady-to-use355-5309 100 ml x 6 bottles355-5307 200 ml x 6 bottles
Dehydrated356-4035 500 g
Tryptose Lauryl Sulfate/BrothDehydrated356-4774 500 g
Tryptone-Salt/DiluentReady-to-use355-5754 9 ml x 25 tubes355-5756 90 ml x 6 bottles355-5796 3 L x 4 bags
Dehydrated356-4544 500 g
44°C18-24 hrs
* Please refer to ISO 7218: 2007
(2 X)*
Escherichia coli - ISO standards
Room t° 15 minand 37°C 4 hrs ± 1 hr
44°C18-24 hrs
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Food Safety and Quality Diagnostics • Bio-Rad Solutions | | 5
Routine Testing • Quality Indicator
ISO 7251 (07/2005)
General guidance for detection and enumeration
of presumptive E. coli
Most probable number technique
ISO/TS 16649-3 (09/2005)
Horizontal method for the enumerationof β-glucuronidase-positive E. coli
Part 3: Most probable numbertechnique using 5-bromo-4-chloro-
3-indolyl -D-glucuronide
Sample/initial suspension
RAPID’E.coli 2
Dilutions
Tryptose LaurylSulfate
EC Medium
Peptone Water
Indole test reading and calculation
Sample/initial suspension
Singleconcentration
Doubleconcentration
37°C24-48 hrs ± 2 hrs
37°C24-48 hrs ± 2 hrs
44°C24-48 hrs± 2 hrs
44°C/37°C18-24 hrs
Direct enumerationof typical colonies*
* Enumeration of typical colonies for E. coli at 44°C; for E. coli and coliforms at 37°C
** Please refer to AOAC certificate for validated matrices
Sample/initial suspension
* For detection method only
(or)*
44°C48 hrs± 2 hrs
Each tube showing the presence of blue colonies on TBX
should be considered positive
Calculate the most probable number from the number of positive tubes
at each dilution
44°C20-24 hrs
37°C24 hrs ± 2 hrs
Add 10 ml in 3 tubes of double
strength MMG
And subsequent dilution Add 1 ml in 3 tubes
of single strength MMG
Subculture each tube presentingyellow coloration on a dish of TBX
Escherichia coli - ISO standards
Escherichia coliAlternative method: RAPID’E.coli 2 Agar
Enumeration of E. coliNF VALIDATION according to ISO 16140N° BRD 07/1-07/93 and BRD 07/7-12/04
AOAC-RI approved N° 050601**NordVal approved N° 020 Health Canada (Pending)
All food stuffs
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Routine Testing • Quality Indicator
6 | | Food Safety and Quality Diagnostics • Bio-Rad Solutions
Coliforms - ISO standards
ISO 4831 (08/2006)
Microbiology of food and animal feeding stuffs Horizontal method for the detection
and enumeration of coliforms
MPN technique
Sample
Tryptose Lauryl Sulfate
Dilutions
30°C or 37°C24-48 hrs ± 2 hrs
30°C or 37°C24 hrs ± 2 hrs
30-37°C24 hrs (+ 24 hrs)
Reading (gas +)
BGBLB
Single concentration
Double concentration(or)*
* For detection method only
Ordering Information
Catalog # Description
BGBLB (Brilliant Green BileLactose Broth)/BrothReady-to-use357-8024 10 ml x 25 tubes
Buffered Peptone Water/BrothReady-to-use355-4179 225 ml x 6 bottles355-5795 3 L x 4 bags355-5790 5 L x 2 bags
Dehydrated356-4684 500 g
RAPID’E.coli 2/AgarReady-to-use355-5297 200 ml x 6 bottles355-5299 100 ml x 6 bottles
Dehydrated356-4024 500 g
Tryptose Lauryl Sulfate/BrothDehydrated356-4774 500 g
Tryptone-Salt/DiluentReady-to-use355-5754 9 ml x 25 tubes355-5756 90 ml x 6 bottles355-5796 3 L x 4 bags
Dehydrated356-4544 500 g
VRBL (Violet Red Bile Lactose)/AgarReady-to-use355-4799 100 ml x 6 bottles
Dehydrated356-4594 500 g
VRBL Coliforms
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Food Safety and Quality Diagnostics • Bio-Rad Solutions | | 7
Routine Testing • Quality Indicator
Coliforms - ISO standards
ISO 4832 (02/2006)
Microbiology of food and animalfeeding stuffs
Horizontal methodfor the enumeration of coliforms
Colony-count technique
Enumeration of coliformsNF VALIDATION according to ISO 16140
N° BRD 07/8-12/04AOAC-RI approved N° 050601**
NordVal approved N° 020Heath Canada (Pending)
All food stuffs
Direct enumerationof typical colonies*
Sample/initial suspension
RAPID’E.coli 2
Enumeration of characteristic colonies
and confirmation for atypical colonies
Sample/initial suspension
Dilutions Dilutions
* Enumeration of typical colonies for E. coli at 44°C; for E. coli and coliforms at 37°C
** Please refer to AOAC certificate for validated matrices
VRBL
30-37°C24 hrs ± 2 hrs
37°C18-24 hrs
Coliforms
• Diameter ≥ 0.5 mm• Purple with or without a red
zone of precipitated bile
* Please refer to ISO 7218: 2007
(2 X)*
ColiformsAlternative method: RAPID’E.coli 2 Agar
RAPID’E.coli 2 E. coli Other Coliforms
Total Coliforms
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Routine Testing • Quality Indicator
8 | | Food Safety and Quality Diagnostics • Bio-Rad Solutions
Enterobacteriaceae
ISO 21528-1 (08/2004)
Microbiology of food and animal feeding stuffs Horizontal methods for the detection
and enumeration of Enterobacteriaceae
Part 1: Detection and enumeration by MPN technique with pre-enrichment
VRBG
Oxidase Dextrose BCP
Yellow color (+)
EE
Biochemical confirmation
ConfirmationNutrient (2.8%)
Sample
Buffered Peptone Water
37°C24 hrs ± 2 hrs
Ordering Information
Catalog # Description
Buffered Peptone Water/BrothReady-to-use355-4179 225 ml x 6 bottles355-5795 3 L x 4 bags355-5790 5 L x 2 bags
Dehydrated356-4684 500 g
EE/Broth Dehydrated356-4794 500 g
Nutrient (2.8%)/AgarReady-to-use355-4478 8 ml x 25 tubes
Dehydrated356-4484 500 g
Oxidase Test355-3834 2 x vials of 50 oxidase disks
VRBG (Violet Red Bile Glucose)/AgarReady-to-use355-4239 100 ml x 6 bottlesDehydrated356-4584 500 g
Non-selective pre-enrichment
37°C18 hrs ± 2 hrs
Selective enrichment
37°C24 hrs ± 2 hrs
37°C24 hrs ± 2 hrs
37°C24 hrs ± 2 hrs
VRBG Enterobacteriaceae
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Routine Testing • Quality Indicator
Enterobacteriaceae
ISO 21528-2 (08/2004)
Microbiology of food and animal feeding stuffs
Horizontal methods for the detection and enumeration of Enterobacteriaceae
Part 2: Colony-count technique
Reading
Oxidase (-) Dextrose BCP
Yellow color (+)
Biochemical confirmation
Confirmation Nutrient (2.8%)
Sample/initial suspension
VRBG
* Please refer to ISO 7218: 2007
Dilutions
(2 X)*
37°C24 hrs± 2 hrs
37°C24 hrs± 2 hrs
37°C24 hrs± 2 hrs
Enumeration of Enterobacteriaceae
NF VALIDATION according to ISO 16140
N° BRD 07124-11/13
Direct enumeration*
Sample/initial suspension
RAPID’Enterobacteriaceae
Dilutions
* Suitable with an automatic plate counter** Alternatively, a temperature of 30°C or
35°C can be chosen
37°C**± 1°C24 hrs± 2 hrs
EnterobacteriaceaeAlternative method: RAPID’Enterobacteriaceae
RAPID’Enterobacteriaceae
Enterobacteriaceae
Non Enterobacteriaceae
Ordering Information
Catalog # Description
RAPID’Enterobacteriaceae/AgarReady-to-use355-4012 200 ml x 6 bottles
Dehydrated356-4004 500 g
Tryptone-Salt/DiluentReady-to-use355-5754 9 ml x 25 tubes355-5756 90 ml x 6 bottles355-5796 3 L x 4 bags
Dehydrated356-4544 500 g
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Pathogen Testing
10 | | Food Safety and Quality Diagnostics • Bio-Rad Solutions
Salmonella - ISO standard
EN ISO 6579/A1 (02/2006)
Microbiology of food and animal feeding stuffsHorizontal method for the detection of Salmonella spp.
Amendment 1 Appendix D: Detection of Salmonella spp. in animal feces and in samplesfrom primary production stages
Buffered Peptone Water
Biochemical and serological confirmationTSI Agar and other test
37°C ± 1°C24 hrs ± 3 hrs
ConfirmationNutrient 2.1%
Sample
XLD RAPID’Salmonellaor XLT 4
or Kristensenor Hektoen
Non-selectivepre-enrichment
37°C ± 1°C18 hrs ± 2 hrs
Appendix D ProtocolSelectiveenrichment
37°C ± 1°C24 hrs ± 3 hrs41.5°C ± 1°C
24 hrs ± 3 hrs
RappaportVassiliadis Soya
MKTTn
Plating-out
MSRV
41.5°C ± 1.5°C24 hrs ± 3 hrs(+24 hrs for negative)
Selectiveisolation
37°C ± 1°C24 hrs ± 3 hrs
Confirmation
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Pathogen Testing
FDA-BAM Salmonella protocol FSIS Salmonella MLG 4.07
Lactose Brothor Trypticase Soy Broth with/without Ferrous Sulfateor Universal Pre-enrichment Broth with/without Ferric
Ammonium Citrateor Buffered Peptone Water
Buffered Peptone Wateror mTSB (for raw beef, carcass, and environmental samples)
Biochemical and serological confirmationTSI and LIA Agar
Biochemical and serological confirmationTSI and LIA Agar
Confirmation
Sample Sample
XLD Hektoen Bismuth Sulfite
Brillant Green Sulfat Agar
and XLT4 or DMLIA
Non-selectivepre-enrichmentPre-enrichment depends on the matrix
Non-selectivepre-enrichment
Plating-out
35°C or 43°C24 hrs ± 2 hrs
42°C24 hrs ± 2 hrs
Selectiveisolation
Plating-out
42°C22-24 hrs
Selectiveisolation
37°C ± 1°C24 hrs ± 3 hrs
Confirmation
35°C18-24 hrs
Selectiveenrichment
37°C24 hrs ± 2 hrs
Selectiveenrichment
37°C18-24 hrsDepending on the food
RappaportVassiliadis Soya
andTretiationate
Broth
Modified Rappaport
Vassiliadis Brothand
TetrathionateBroth
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Pathogen Testing
12 | | Food Safety and Quality Diagnostics • Bio-Rad Solutions
Salmonella - Alternative method: RAPID’Salmonella Agar double enrichment protocol
Salmonella - Alternative method: RAPID’Salmonella Agar short protocol
Salmonella - Alternative method: iQ-Check Salmonella II real-time PCR Kit
BufferedPeptone Water
RappaportVassiliadis Soya
RAPID’SalmonellaRAPID’Salmonella
Sample Sample
Reading & confirmationStandard tests
or Oxidase/OMNI-O/ONPG or iQ-Check Salmonella II
Detection of Salmonella spp.NF VALIDATION according to
ISO 16140N° BRD 07/11-12/05
AOAC-RI approved N° 0507011*All food stuffs and feed
For more information on Bio-Rad’s real-time PCR kits and instruments, please contact your local Bio-Rad sales representative.1 - For AOAC-RI protocol, please contact your local Bio-Rad sales representative2 - 8-hr enrichment protocol available for raw meat (AOAC and AFNOR validated)
37°C18 hrs± 2 hrs
Non-selectivepre-enrichment
41.5°C± 1°C18 hrs± 2 hrs
Selectiveenrichment
37°C24 hrs± 2 hrs
Selectiveisolation
37°C±1°C24 hrs± 2 hrs
Selectiveisolation
41.5°C6-26 hrs
Selectiveenrichment
Plating-out
Plating-out
Reading & confirmationSalmonella Latex or Oxidase/
OMNI-O/ONPGor iQ-Check Salmonella
Detection of Salmonella spp. Certification by NF VALIDATION
according to ISO 16140 N° 07/11-12/05
AOAC-RI approved N° 050701NordVal approved N° 032
Buffered Peptone Water+
Salmonella Capsule Supplement
Buffered Peptone Water
Sample
Detection of Salmonella spp.NF VALIDATION according to ISO 16140
N° BRD 07/06-07/04AOAC-RI approved N° 010803*
NordVal approved N° 038Health Canada approved N° MFLP-38All foods stuffs1,2, feed, and environment
including primary production
Bacterial Lysis & DNA Extraction(Standard or Easy)
Real-time PCR amplification and detection
with iQ-Check Salmonella II
Automatic result analysis and interpretation
Confirmation of positive with RAPID’Salmonella
+/-
37°CFrom 10 hrs ± 2 hrsto 21 hrs ± 1 hr
* Please refer to AOAC certificate for validated matrices
* Please refer to AOAC certificate for validated matrices
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Pathogen Testing
Novobiocin/SupplementDehydrated356-4610 1 g
Nutrient (2.1%)/AgarDehydrated356-4485 500 g
ONPG Test355-3822 Set of 50 disks
Oxidase Test355-3834 2 x vials of 50 oxidase disks
RAPID’Salmonella/Agar Ready-to-use356-3961 90 mm x 20 dishes356-3963 90 mm x 120 dishes
Dehydrated356-4705 500 g
RAPID´Salmonella Capsules/Supplement 356-4709 100 x quantity for 2.5 L356-4710 100 x quantity for 250 ml356-4712 1 x quantity for
100 analyses
RVS/Broth (Rappaport Vassiliadis Soy)Ready-to-use355-5773 10 ml x 25 tubes
Dehydrated356-4324 500 g
Salmonella Latex Kit
Detection of groups B to E and G355-6710 75 tests
Ordering Information
Catalog # Description
Antiserum SalmonellaOmnivalent OMNI-O (A60)356-0781 3 ml dropper bottle
Brilliant Green (Kristensen)/AgarDehydrated356-4464 500 g
Buffered Peptone Water/Broth Ready-to-use355-4179 225 ml x 6 bottles355-5795 3 L x 4 bags355-5790 5 L x 2 bags
Dehydrated356-4684 500 g
Hektoen/Agar Ready-to-use355-4386 100 ml x 6 bottles356-3894 90 mm x 20 dishes
Dehydrated356-4284 500 g
iQ-Check Salmonella II Kit357-8123 96 tests
MKTTN (Muller Kauffmann Tetrathionate Novobiocin)/Broth Ready-to-use355-6140 10 ml x 50 tubes
Dehydrated Base356-4714 500 g
MSRV (Semi-Solid Rappaport Vassiliadis Medium)
Ready-to-use355-6139 200 ml x 6 bottles
Dehydrated356-4325 500 g
Salmonella Confirm Latex Kit355-6711 50 tests
Selenite-Cystine/BrothComplete Medium355-5746 10 ml x 25 tubes
TSB/BrothReady-to-use(containing novobiocin)355-5426 225 ml x 6 bottles
Dehydrated Base356-4426 500 g
TSI (Triple Sugar Iron)/AgarDehydrated356-4384 500 g
XLD (Xylose Lysine Desoxycholate)/AgarReady-to-use354-1751 90 mm x 20 dishes
Dehydrated356-9124 500 g
XLT 4 (Xylose Lysine Tergitol 4)/AgarReady-to-use356-3654 90 mm x 20 dishes
RAPID’Salmonella
Salmonella spp.
Other Enterobacteriaceae
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Pathogen Testing
14 | | Food Safety and Quality Diagnostics • Bio-Rad Solutions
Ordering Information
Catalog # Description
Buffered Peptone Water/Broth Ready-to-use355-4179 225 ml x 6 bottles355-5795 3 L x 4 bags355-5790 5 L x 2 bags
Dehydrated356-4684 500 g
iQ-Check Cronobacter spp. 357-8137 96 tests
RAPID’Sakazakii/AgarReady-to-use356-3971 90 mm x 20 dishes
Dehydrated356-4976 500 g
Trypto-Casein-Soy (TSA)/Agar Ready-to-use 356-3884 90 mm x 20 dishes355-5947 200 ml x 6 bottles 355-5944 8 ml x 25 inclined tubes
RAPID’Sakazakii
Cronobacter spp.
Other Enterobacteriaceae
Cronobacter spp. - ISO standardFDA BAM Cronobacter spp. protocol
ISO/TS 22964:2006Milk and milk products -
Detection of Enterobacter sakazakii
Streak onto RAPID’Sakazakii
1:10Buffered Peptone Water
Perform identification tests on yellow colonies
Typical colonies from each plate are confirmed with real-time PCR
and other techniques
Sample preparationWeigh z g of the test portion
in 9 x z ml Buffered Peptone Water
Samples
Selective enrichmentTransfer 0.1 ml in 10 ml
of mLST/vancomycin medium
Transfer 1 to 5 colonies on TSA
Centrifuge 3,000 x g for 10 min. Decant supernatant
Resuspend pellet in 200 µl PBS
37°C18 hrs ± 2 hrs
44°C24 hrs ± 2 hrs
44°C24 hrs ± 2 hrs
25°C48 hrs
36°C ± 1°C24 hrs ± 2 hrs
Incubation
40 ml
Spread and streak plates
100 µl
PCR PCRDFI
AgarR&F Agar
36°C ± 1°C18-24 hrs
Incubation
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Food Safety and Quality Diagnostics • Bio-Rad Solutions | | 15
Pathogen Testing
Cronobacter spp. - Alternative method: RAPID’Sakazakii Agar short protocol
RAPID’Sakazakii short protocolNF VALIDATION according to
ISO 16140N° BRD 07/22-05/12
Streak onto RAPID’Sakazakii
Perform identification tests on yellow colonies
Sample preparationWeigh z g of the test portion
in 9 x z ml BPW
Transfer 1 to 5 colonies on TSA
37°C18 hrs ± 2 hrs
44°C24 hrs ± 2 hrs
25°C48-72 hrs
Cronobacter spp. - Alternative method: iQ-Check Cronobacter spp. real-time PCR Kit
Buffered Peptone WaterSuppl. Buffered Peptone Water
Buffered Peptone Water
Environmental sample
Detection of Cronobacter spp. Infant Formulas and environment
NF VALIDATION pending
Bacterial Lysis & DNA Extraction(Standard or Easy)
Real-time PCR amplification and detection with iQ-Check Cronobacter spp.
Automatic result analysis and interpretation
Confirmation of positive with RAPID’Sakazakii
+/-
37°C18 hrs ± 2 hrs
Infant formula
37°C20 hrs ± 2 hrs
37°C4 hrs ± 1 hr
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Pathogen Testing
16 | | Food Safety and Quality Diagnostics • Bio-Rad Solutions
Listeria monocytogenes - ISO standards
EN ISO 11290-1/A1 (02/2005)
Horizontal method for the detection and enumeration of L. monocytogenes
Part 1: Detection method
Reading andconfirmation
Isolation
Primary enrichmentIsolation Secondary enrichment
Sample
Fraser ½
Fraser 1(0.1 ml culture/10 ml)
Listeria spp.
- TSYEA agar- Catalase +- Gram test- Mobility
L.monocytogenes
- TSYEB- Hemolysis- Carbohydrates- CAMP test
Agar Listeria accordingto Ottaviani and Agosti
Other solid selective medium
30°C24 hrs ± 2 hrs
35°C or 37°C48 hrs ± 2 hrs
37°C ± 1°C24 hrs ± 3 hrs(+28 hrs ± 2 hrs)*
30°C-35°C-37°C(dependingof the media)24-48 hrs ± 2 hrs
Reading andconfirmation
37°C ± 1°C24 hrs ± 3 hrs(+24 hrs ± 3 hrs)
30°C-35°C-37°C(depending onthe media)24-48 hrs ± 2 hrs
Agar Listeria according
to Ottaviani and Agosti
Other solid selective medium
* If low or absent colony growth observed after 24 hours.
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Pathogen Testing
Listeria monocytogenes - ISO standardsOrdering InformationCatalog # Description
AL (Agar Listeria accordingto Ottaviani and Agosti)/Agar Ready-to-use356-3695 90 mm x 20 dishes356-3965 90 mm x 120 dishes
Dehydrated Base356-4043 500 g
Base in vials (to be supplemented)355-5200 237.5 ml x 6 bottles
Supplement 1 356-4201 100 capsules (1 capsule qsp 250 ml)356-4041 10 vials
Supplement 2356-4042 10 vials
Buffered Peptone Water/BrothReady-to-use355-4179 225 ml x 6 bottles355-5795 3 L x 4 bags355-5790 5 L x 2 bags
Dehydrated356-4684 500 g
Fraser ½/BrothComplete Medium355-5797 225 ml x 6 bottles355-5794 3 L x 4 bags355-5792 5 L x 2 bags
Dehydrated Base356-4604 500 g
Supplement356-4616 10 vials
Fraser 1/Broth Complete Medium355-4569 10 ml x 25 tubes
Dehydrated Base356-4604 500 g
Supplement356-4615 10 vials
Oxford/AgarReady-to-use356-3664 90 mm x 20 dishes
Palcam/AgarDehydrated Base356-4754 500 g
Supplement356-4752 10 vials
EN ISO 11290-2/A1 (02/2005)
Horizontal method for the detection and enumeration of L. monocytogenes
Part 2: Enumeration method
Listeria spp.
- TSYEA agar- Catalase +- Gram test- Mobility
Listeriamonocytogenes
- TSYEB- Hemolysis- Carbohydrates- CAMP test
Enumeration and confirmation tests
Sample
Fraser ½ without LiCland selective supplement
or Buffered Peptone Water
Agar Listeria accordingto Ottaviani and Agosti
Dilution
20°C ± 2°C1 hr ± 5 min
37°C24 hrs ± 3 hrs(+ 24 hrs ± 3 hrs)
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Pathogen Testing
18 | | Food Safety and Quality Diagnostics • Bio-Rad Solutions
FDA-BAM Listeria monocytogenes protocol
BLED
+ supplements
Sample
37°C24 hrs (+24 hrs)
37°C24 hrs (+24 hrs)
30°C4 hrs
30°C24 hrs ± 2 hrs (48 hrs ± 2 hrs)*
Oxford or
Modified Oxford (MOX)
PalcamRAPID’L.mono
Agar Listeria according to Ottaviani and
Agosti
or and or
Reading, isolation, and confirmation
Listeria spp.
- TSYEA agar- Catalase +- Gram test- Mobility
L.monocytogenes
- TSYEB- Hemolysis- Carbohydrates- CAMP test
or
* If low or absent colony growth observed after 24 hours
Reading and confirmation
Sample
LEB
Health Canada Listeria monocytogenes protocol MFHPB-30
30°C24 hrs
30°C24 hrs
30°C48 hrs
0.1 ml MFB 0.1 ml MFB35°C24 hrs(48 hrs)
35°C24 hrs(48 hrs)
Oxford + RAPID’L.mono
Oxford + RAPID’L.mono
35°C24 – 48 hrs
35°C24 – 48 hrs
Oxford + RAPID’L.mono
35°C24 – 48 hrs
- Motility- Hemolysis- Mannitol, rhamnose
and xylose (or other equivalent compendium-approved methods)
- Real-Time PCR Amplification
- Catalase- Gram test- Serology
and (optional)
Real-timePCRamplification
Real-timePCRamplification
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Food Safety and Quality Diagnostics • Bio-Rad Solutions | | 19
Pathogen Testing
Listeria monocytogenes - Alternative method: RAPID’L.mono Agar
Detection of Listeria monocytogenesNF VALIDATION according to ISO 16140
N° BRD 07/4-09/98AOAC-RI approved N° 0304061*
NordVal approved All food stuffs and environment
30°C24 hrs± 2 hrs
37°C24 hrs± 2 hrs
Isolation onRAPID’L.mono
Reading andconfirmation tests
Fraser ½ Broth
Sample
Standard Tests
- TSYEB- Hemolysis- Carbohydrates- CAMP test
iQ-Check Listeria monocytogenes II
Spot on Agar Listeria according to Ottaviani and Agosti
Rhamnose Test
or oror
For more information on Bio-Rad’s real-time PCR kits and instruments, please contact your local Bio-Rad sales representative.1 - For AOAC-RI protocol, please contact your local Bio-Rad sales representative
Enumeration of Listeria monocytogenesNF VALIDATION according to
ISO 16140N° BRD 07/05-09/01
All food stuffs and environment
37°C24 hrs± 2 hrs
Isolation onRAPID’L.mono
Directenumeration
Buffered Peptone Water
Sample
20°C1 hr
Resuscitation
For a NF VALIDATION validated enumeration method, in the event of a positive result using RAPID’L.mono for enumeration, confirmation is not necessary when the presence of Listeria monocytogenes has been confirmed during detection.* Please refer to AOAC certificate for validated matrices
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Pathogen Testing
20 | | Food Safety and Quality Diagnostics • Bio-Rad Solutions
Listeria monocytogenes - Alternative method: iQ-Check Listeria monocytogenes II real-time PCR Kit
(1) Listeria Special Broth (LSB) or (2) Fraser ½ Broth
Sample
Detection of Listeria monocytogenesNF VALIDATION according to ISO 16140
N° BRD 07/10-04/05AOAC-RI approved N° 010802*
NordVal approved N° 037All food stuffs and environment
Bacterial Lysis & DNA ExtractionStandard (Fraser ½ and LSB) or Easy (LSB)
Real-time PCR amplification and detection with iQ-Check Listeria monocytogenes II
Automatic result analysis and interpretation
Confirmation of positive with RAPID’L.mono
+/-
30°C23 hrs ± 1 hr (1)
or 25 hrs ± 1 hr (2)
RAPID’L.mono
L. mono
L. ivanovii
L. innocua
L. welshimeri
Ordering InformationCatalog # Description
Buffered Peptone Water/Broth Ready-to-use355-4179 225 ml x 6 bottles355-5795 3 L x 4 bags355-5790 5 L x 2 bags
Dehydrated356-4684 500 g
iQ-Check Listeria monocytogenes II Kit357-8124 96 tests
LSB (Listeria Special Broth)Ready-to-use355-5703 225 ml x 6 bottles355-5793 5 L x 2 bags
Dehydrated356-4703 500 g356-4753 5 kg
RAPID’L.mono/AgarReady-to-use356-3964 90 mm x 120 dishes356-3694 90 mm x 20 dishes355-5294 Kit: 190 ml agar + 2 suppl.
Dehydrated Base356-4293 500 g
Supplement 1356-4294 10 vials
Supplement 2356-4746 10 vials
Rhamnose Test355-3669 28 tests
* Please refer to AOAC certificate for validated matrices
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Food Safety and Quality Diagnostics • Bio-Rad Solutions | | 21
Pathogen Testing
Listeria spp. Alternative method: RAPID’L.mono Agar
Detection of Listeria spp.NF VALIDATION according to
ISO 16140N° BRD 07/4-09/98
All food stuffs and environment
37°C(24 hrs) 48 hrs
Isolation onRAPID’L.mono
Reading and confirmation tests
Fraser ½ Broth
Sample
30°C24 hrs± 2 hrs
Standard Tests- TSYEA- Gram test- Catalase
iQ-Check Listeria spp.
or
Listeria spp. - Alternative method: RAPID’Listeria spp. Agar
Detection of Listeria spp.NF VALIDATION according to ISO 16140
N° BRD 07/12-12/06AOAC-RI approved N° 080701*All food stuffs and environment
30°C24 hrs ± 2 hrs
37°C24 hrs ± 2 hrs
Isolation onRAPID’Listeria spp.
Reading andconfirmation tests
Fraser ½ Broth or UVM
Sample
Standard Tests
- Gram test- Catalase
Spot on Palcam
Spot on RAPID’L.mono + Gram test+ Catalase
iQ-Check Listeria spp.
or oror
* Please refer to AOAC certificate for validated matrices
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Pathogen Testing
22 | | Food Safety and Quality Diagnostics • Bio-Rad Solutions
Ordering Information
Catalog # Description
Fraser ½/BrothReady-to-use355-5797 225 ml x 6 bottles355-5794 3 L x 4 bags355-5792 5 L x 2 bags
Dehydrated Base356-4604 500 g
Supplement356-4616 10 vials
iQ-Check Listeria spp. Kit357-8113 96 tests
LSB (Listeria Special Broth)Ready-to-use355-5703 225 ml x 6 bottles355-5793 5 L x 2 bags
Dehydrated356-4703 500 g356-4753 5 kg
RAPID’Listeria spp./AgarDehydrated Base356-4744 500 g
Supplement 1356-4745 10 vials
Supplement 2
356-4746 10 vials
RAPID’L.mono/AgarDehydrated Base356-4293 500 g
Supplement 1356-4294 10 vials (1 vial qsp 500 ml)
Supplement 2356-4746 10 vials (1 vial qsp 500 ml)
For more information on Bio-Rad’s real-time PCR kits and instruments, please contact your local Bio-Rad sales representative.
* For AOAC-RI protocol, please contact your local Bio-Rad sales representative.
Listeria spp. - Alternative method: iQ-Check Listeria spp. real-time PCR Kit
Listeria Special Broth (LSB)
Sample
Detection of Listeria spp.NF VALIDATION according to ISO 16140
N° BRD 07/13-05/07AOAC-RI approved N° 090701*
Health Canada Validation - MFLP-39All food stuffs and environment
Bacterial Lysis & DNA Extraction((1) Standard or (2) Easy)
Real-time PCR amplification and detection with iQ-Check Listeria spp.
Automatic result analysis and interpretation
Confirmation of positive with RAPID’Listeria spp. or RAPID’L.mono
+/-
30°C23 hrs ± 1 hr (1)
or 25 hrs ± 1 hr (2)
* Please refer to AOAC and Health Canada certificates for validated matrices
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Food Safety and Quality Diagnostics • Bio-Rad Solutions | | 23
Pathogen Testing
Ordering Information
Catalog # Description
Buffered Peptone Water/Broth Ready-to-use355-4179 225 ml x 6 bottles355-5795 3 L x 4 bags355-5790 5 L x 2 bags
Dehydrated356-4684 500 g
iQ-Check E.coli O157:H7 Kit357-8114 96 tests
iQ-Check STEC SerO357-8140 32 tests
iQ-Check STEC VirX357-8139 96 tests
mTSB/BrothReady-to-use (containing novobiocin)355-5426 225 ml x 6 bottles
Base356-4426 500 g
Novobiocin/SupplementDehydrated356-4610 1 g
RAPID’E.coli O157:H7/Agar Base356-4748 100 g
SEB (STEC Enrichment Broth)/Broth356-4001 500 g356-4002 5 kg
For more information on Bio-Rad’s real-time PCR kits and instruments, please contact your local Bio-Rad sales representative.
RAPID’E.coli O157:H7 E. coli O157:H7 Other E. coli
Escherichia coli O157:H7 - Alternative method: RAPID’E.coli O157:H7 Agar
Detection of Escherichia coli O157:H7NF VALIDATION according to ISO 16140
N° BRD 07/14-09/07AOAC-RI approved N° 060701*All food stuffs and environment
Immunomagnetic-separation
Reading and confirmation testsStandard tests or Latex or
iQ-Check E.coli O157:H7
mTSB + Novobiocin
Isolation on RAPID’E.coli O157:H7
Sample
41°C16-24 hrs
37°C24 hrs ± 2 hrs
RAPID’E.coli O157:H7 can also be used as a second medium in the ISO 16654 standard* Please refer to AOAC certificate for validated matrices
* For AOAC-RI protocol, please contact your local Bio-Rad sales representative.
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Pathogen Testing
24 | | Food Safety and Quality Diagnostics • Bio-Rad Solutions
Escherichia coli O157:H7 - Alternative method: iQ-Check E.coli O157:H7 real-time PCR Kit
Buffered Peptone Water
Sample
Detection of Escherichia coli O157:H7NF VALIDATION according to ISO 16140 N° BRD 07/15-06/08
Raw beefAOAC-RI approved N° 020801*
Raw ground beef, fresh spinash, apple cider
Bacterial Lysis &Easy DNA Extraction
Real-time PCR amplification and detection with iQ-Check E.coli O157:H7
Result analysis and interpretation
Confirmation of posiitve with RAPID’E.coli O157:H7
+/-
41.5°C8-24 hrs
STEC iQ-Check STEC VirX and SerO real-time PCR Kits
SEB enrichment
Sample
Detection and Isolation of non-O157 Shiga Toxin-Producing
E. coli (STEC) from ground beef
USDA No Objection Letter
Bacterial Lysis & DNA Extraction
Real-time PCR amplification and detection with
iQ-Check STEC VirX (Automatic results analysis and
interpretation)
Real-time PCR amplification and detection with
iQ-Check STEC SerO (Automatic results analysis and
interpretation)
+/-
41.5°C10-15 hrs
* For AOAC-RI protocol, please contact your local Bio-Rad sales representative.
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Food Safety and Quality Diagnostics • Bio-Rad Solutions | | 25
Pathogen Testing
Campylobacter spp. - ISO standard
ISO 10272-1:2006
Microbiology of food and animal feeding stuffsHorizontal method for detection and enumeration
of Campylobacter spp.
Part 1: Detection method
Sample
41.5°C44 ± 4 hrs
37°C 4-6 hrs
+
Bolton Broth
(microaerobic atmosphere)
41.5°C44 ± 4 hrs
(microaerobic atmosphere)
41.5°C24-48 hrs
Innoculate a steril loop
mCDD Agar Other solid selective medium
Isolation Columbia blood Agar
Reading and confirmation
Study of growth Columbia blood Agar
Study of growth Columbia blood Agar
Oxidase
25°C44 ± 4 hrs
(microaerobic atmosphere)
41.5°C44 ± 4 hrs
(aerobic atmosphere)
Examination of morphology and motility
ISO/TS 10272-2:2006
Microbiology of food and animal feeding stuffsHorizontal method for detection and enumeration
of Campylobacter spp.
Part 2: Enumeration method
Sample
Isolation 0.1 mlmCDD (x2)
41.5°C24-48 hrs
Isolation Columbia blood Agar
Reading and confirmation
Study of growth Columbia blood Agar
Study of growth Columbia blood Agar
Oxidase
25°C44 ± 4 hrs
(microaerobic atmosphere)
41.5°C44 ± 4 hrs
(aerobic atmosphere)
Examination of morphology and motility
Dilution
41.5°C40-48 hrs
(microaerobic atmosphere)
Select 5 colonies
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Pathogen Testing
26 | | Food Safety and Quality Diagnostics • Bio-Rad Solutions
FDA-BAM Campylobacter spp. protocol
Rinse gently; shake 5 min. (25 rpm on bench-top shaker)Remove net-lined bag
100 ml Bolton Broth with antibiotics in net-lined bag
Preenrichment
Most samples(except: shellfish, milk, frozen dairy products)
25 g sample (50 g vegetables)
Streak 24 and 48 hrs enrichments on isolation agar
37°C4 hrs
37°C4 hrs
Microaerobic MicroaerobicExcept: bubbling at 30°C)
or
Enrichment42°C20-44 hrs
Microaerobic
Identification
Abeyta-Hunt-Bark agar or mCCDA
37°C or 42°C*24-72 hrs* in microaerobic
Incubate
Campylobacter spp. - Alternative method: iQ-Check Campylobacter spp. real-time PCR Kit
Detection of Campylobacter jejuni, Campylobacter coli and Campylobacter lari
AOAC-RI approved N° 031209*
For chicken carcass rinses, turkey swabs and raw ground chicken
Bacterial Lysis and DNA Extraction(Easy)
Real-time PCR amplification and detection with iQ-Check Campylobacter
Result analysis and interpretation
+/-
41.5°C ± 1°C20 – 24 hrs
37°C ± 1°C4 – 6 hrs
42°C 24 hrs
Suppl. Bolton Broth 2X BF-BEB
Food Sample Carcass rinse or sponge
+
* For AOAC-RI protocol, please contact your local Bio-Rad sales representative.
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Food Safety and Quality Diagnostics • Bio-Rad Solutions | | 27
Pathogen Testing
Campylobacter spp. - Alternative method: RAPID’Campylobacter spp.
Sample enrichment Suspension/Dilution
Ordering Information
Catalog # Description
Buffered Peptone Water/BrothReady-to-use355-4179 225 ml x 6 bottles355-5795 3 L x 4 bags355-5790 5 L x 2 bags
Dehydrated356-4684 500 g
Confirmation356-3784 Columbia + Sheep
Blood/Agar, 90 mm x 20 dishes
355-4164 Physiologic Sterile Water (0.85%), 9 ml x 25 tubes
355-3834 Oxidase test, 2 x 50 disks
355-4154 Sterile Distilled Water, 9 ml x 25 tubes
356-4297 Campylobacter Confirm Latex kit, 50 tests
iQ-Check Campylobacter/Kit357-8135 96 tests
RAPID’Campylobacter/AgarDehydrated Base356-4295 500 g
Supplement356-4296 10 vials (1 vial qsp 400 ml)
Tryptone Salt/DiluentReady-to-use355-5756 90 ml x 6 bottles355-5754 9 ml x 25 tubes355-5747 9 ml x 100 tubes355-5796 3 L x 4 bags
Dehydrated356-4544 500 g
Preston Broth
Reading and confirmation testsCampylobacter Confirm Latex
oriQ-Check Campylobacter
orStandard tests
Reading and confirmation testsCampylobacter Confirm Latex
oriQ-Check Campylobacter
orStandard tests
41.5 °C ± 1°C24 hrs ± 2 hrs
Detection ofCampylobacter spp.
Enumeration ofCampylobacter spp.
NF VALIDATION according to ISO 16140 N° BRD 07/25–01/14
Isolation on RAPID’Campylobacter
Isolation on RAPID’Campylobacter
41.5 °C ± 1°C24-48 hrs
(microaerobic atmosphere)
41.5 °C ± 1°C(24 hrs) 48 hrs
(microaerobic atmosphere)
RAPID’Campylobacter Campylobacter spp.
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Pathogen Testing
28 | | Food Safety and Quality Diagnostics • Bio-Rad Solutions
Staphylococcus aureus - ISO standards
NF EN ISO 6888-2/A1 (12/2003)
Horizontal method for the enumeration of coagulase-positive
Staphylococci (S. aureus and species)
Technique using rabbit plasma fibrinogen agar medium
NF EN ISO 6888-3 (06/2003)
Horizontal method for the enumeration wof coagulase-positive
Staphylococci (S. aureus and species)
Detection and MPN techniquefor low numbers
Direct enumeration of typical coagulase-positive Staphylococci
Sample/initial suspensionand following dilution
Rabbit Plasma
Reading
Confirmation of typical colonieson BHI
Directenumerationof typicalcoagulase-positive colonies
Sample/initial suspension
NF EN ISO 6888-1/A1 (01/2004)
Horizontal method for the enumeration of coagulase-positive
Staphylococci (S. aureus and other species)
Technique including confirmation of colonies
ConfirmationBrain-Heart Infusion
Rabbit Plasma
Coagulation reading
Sample/initial suspensionand following dilution
Enumeration
Baird-ParkerBaird-Parker
for RPF+ RPF sup.
Baird-Parker Baird-Parkerfor RPF
+ RPF sup.or
Giolitti and CantoniBroth
Simple concentration
Giolitti and CantoniBroth
Double concentration(or)*
37°C24 hrs ± 2 hrs (+24 hrs ± 2 hrs)*
35°C or 37°C18-24 hrsand 48 hrs
35°C or 37°C4-6 hrs or 24 hrs
35°C or 37°C24 hrs ± 2 hrs(+ 24 hrs ± 2 hrs)**
35°C or 37°C24 hrs ± 2 hrs
(2 X)*
Typical Staphylococci grey to black colonies
with a surrounding clear halo and a white opaque zone
* Please refer to ISO 7218: 2007** If low or absent colony growth observed
after 24 hours. * Please refer to ISO 7218: 2007
* For detection method only* If low or absent colony growth observed
after 24 hours.
(2 X)*
37°C24-48 hrs
37°C24-48 hrs
37°C24 hrs
37°C4-6 hrsor 24 hrs
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Food Safety and Quality Diagnostics • Bio-Rad Solutions | | 29
Pathogen Testing
Staphylococcus aureus Alternative method: RAPID’Staph Agar
Enumeration of Coagulase-positive Staphylococci
NF VALIDATION according to ISO 16140N° BRD 07/9-02/05
AOAC-RI approved N° 080602*All food stuffs, feed and environment
FDA BAM Staphylococcus aureus
Sample/initial suspension
Enumeration and Confirmation
Sample
Confirmation
Pastorex™
Staph Plus
Spot on Baird-Parkerfor RPF
or
Ordering Information
Catalog # Description
AdditivesEgg Yolk with Potassium Tellurite355-4201 5 ml x 1 vial355-4205 25 ml x 1 bottle
Sulfamethazine Solution (0.2%)356-2682 2.5 ml x 1 vial
Baird-Parker/Agar Complete Medium356-3991 90 mm x 20 dishes
Base356-4814 500 g
Baird-Parker for RPF/Agar Complete Medium357-8618 90 ml x 6 bottles + 6 vials of suppl.356-3996 90 mm x 20 dishes
Dehydrated Base356-4814 500 g
RPF Supplement356-4618 10 vials
BHI (Brain Heart Infusion)/BrothReady-to-use355-3664 10 ml x 25 tubes
Dehydrated356-4014 500 g
Pastorex™ Staph Plus Latex355-6356 50 tests355-6353 5 x 50 tests
Rabbit PlasmaFreeze-dried355-6352 Pack for 20 reactions
Special broth355-3544 9 ml x 25 tubes
RAPID’Staph/AgarReady-to-use356-3960 90 mm x 20 dishes
Base356-4704 500 g
* For AOAC-RI protocol, please contact your local Bio-Rad sales representative
* Please refer to AOAC certificate for validated matrices
Spread 1 ml on 3 platesBaird Parker
Dilutions
35°C45-48 hrs
RAPID’Staph
37°C24 hrs ± 2 hrs
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Pathogen Testing
30 | | Food Safety and Quality Diagnostics • Bio-Rad Solutions
Clostridium perfringens
NF EN ISO 7937 (02/2005)
Horizontal method for the enumeration of
Clostridium perfringens
Colony-count technique
Sulfite-reducing bacteria
ISO 15213 (05/2003)
Microbiology of food and animal feeding stuffs
Horizontal method for the enumeration of sulfite-reducing
bacteria growing under anaerobic conditions
Reading
Food sample/initial suspension
Dilution
Confirmation
Enumeration of typical black colonies
Confirmation of 5 isolated colonies with
Thioglycolate + Resazurin
Food sample/initial suspension
LactoseSulfite
MobilityNitrate
LactoseGelatine
Clostridiumperfringens(yellow/gas+)
Clostridiumperfringens(red coloration/gas+)
Clostridiumperfringens(black precipitate/
gas+)
37°C18-24 hrs
Anaerobescondition
TSC
TSC (add D-cycloserine)
Ordering Information
Catalog # Description
Lactose Sulfite/BrothBase356-4914 100 g
Sheep Blood/AgarReady to use356-3784 90 mm x 20 dishes
Dehydrated Base356-4674 500 g
Sheep Blood Supplement355-6652 50 ml vial
Thioglycolate + Resazurin/BrothReady to use356-3613 10 ml x 25 tubes
Dehydrated356-4084 500 g
TSC without D-Cycloserine/Agar - Base MediumReady to use355-4419 100 ml x 6 bottles
Base356-9644 500 g
TSC Clostridium perfringens
Heat treatment 20 min 75°C
(2 X)*
Thermophilous bacteria
suspicion50°C 24-48 hrs
37°C24-48 hrsanaerobiccondition
* Please refer to ISO 7218: 2007 * Please refer to ISO 7218: 2007
(2 X)*
Anaerobes condition
37°C20 hrs ± 2 hrs
37°C18-24 hrs
46°C24 hrs
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Food Safety and Quality Diagnostics • Bio-Rad Solutions | | 31
Pathogen Testing
ShigellaBacillus cereus
NF EN ISO 7932 (07/2005)
General guidance for the enumeration of B. cereus
Colony-count technique at 30°C
NF EN ISO 21567 (03/2005)
Horizontal methodfor the detection of Shigella spp.
Confirmation teston 5 well isolated colonies
Food sample/initial suspension
Sheep Blood/Agar
Reading and interpretation
Mossel (MYP)
37°C20-24 hrs(+48 hrs)*
Plating-out
Shigella Broth + NovobiocinpH 7 ± 0.2
Spot on typical colonies on Nutrient (2.1%)
Food sample/initial suspension
Biochemical and serologicalconfirmation
Ordering Information
Catalog # Description
Hektoen/AgarReady-to-use355-4386 100 ml x 6 bottles356-3894 90 mm x 20 dishes
Dehydrated356-4284 500 g
Mac Conkey + Crystal Violet/AgarReady-to-use356-3617 90 mm x 20 dishes
Dehydrated356-9084 500 g
Novobiocin/SupplementDehydrated356-4610 1 g
Nutrient (2.1%)/AgarDehydrated356-4485 500 g
Mossel/Agar (Mannitol Egg Yolk Polymixin)Dehydrated356-9604 500 g
XLD (Xylose Lysine Desoxycholate)/AgarReady-to-use354-1751 90 mm x 20 dishes
Dehydrated356-9124 500 g
Mossel (MYP) Bacillus cereus
* Please refer to ISO 7218: 2007
(2 X)*
30°C18-24 hrs(+24 hrs)
30°C24 hrs ± 2 hrs
41°C16-20 hrs
MacConkey
XLD Hektoen
37°C20-24 hrs
* If low or absent colony growth observed after 24 hours.
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Bio-Rad Solutions
32 | | Food Safety and Quality Diagnostics • Bio-Rad Solutions
PCR MethodsiQ-Check Solutions
Chromogenic MethodsRAPID’ Solutions
Validations
AOACNF
ValidationHealth
CanadaNordVal AOAC
NF Validation
HealthCanada
NordVal
Campylobacter spp.
Coliforms(pending)
Cronobacter spp.
Escherichia coli(pending
Escherichia coli O157:H7
Enterobacteriaceae
Listeria monocytogenes
Detection(pending
Enumeration
Listeria spp.
Salmonella Enteritidis
Salmonella spp.
Salmonella Typhimurium
Staphylococcus aureus
STEC 7 major O-groups*
STEC virulence genes*
* No objection letter delivered from USDA
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Food Safety and Quality Diagnostics • Bio-Rad Solutions | | 33
Bio-Rad Solutions
Meat & Poultry &
Fish Products
Solution SegmentMicroorganism
Water & Wine
Beverages
Dairy Products
Environment Processed Foods
VegetablesProduce
Range for Food Testing
Bacillus cereus
Campylobacter
Clostridium perfringens
Coliforms
Cronobacter sakazakii
Enterobacteriaceae
Escherichia coli
Escherichia coli O157:H7
STEC
Lactobacillus
Listeria spp.
Listeria monocytogenes
Pseudomonas aeruginosa
Salmonella spp.
Salmonella Enteritidis
Salmonella Typhimurium
Staphyloccocus aureus
Yeasts and Molds
RAPID’Chromogenic method iQ-Check PCR method Classical media
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34 | | Food Safety and Quality Diagnostics • Bio-Rad Solutions
14-0686_FSD_Food_Safety_Booklet_Update_17933_FINAL.indd 34 4/10/14 11:14 AM
iQ-Check™ Prep Solution, when the power of automation meets real-time PCR
• Reliable walk-away automation for real-time PCR testing• Designed to fit lab workflow without disrupting it• Increased confidence with improved reproducibility• Optimized to handle all major food pathogens
For more information: foodscience.bio-rad.com
Real-Time PCR Methods • Food Testing
Start, Run, Results
The Bio-Rad PCR Solution, Your Solution
iQ-Check Prep, CFX96 Touch™ deep well and iQ-Check Kits
14-0686_FSD_Food_Safety_Booklet_Update_17933_FINAL.indd 35 4/10/14 11:14 AM
© Bio-Rad 10 17933 14-0686 Rev I 04/2014
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Life ScienceGroup
Bio-Rad Laboratories, Inc.
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