Food Safety and Quality Diagnostics - Bio-Rad · PDF fileFood Testing Food Safety and Quality Diagnostics Bio-Rad Solutions 14-0686_FSD_Food_Safety_Booklet_Update_17933_FINAL.indd

Food Testing

Food Safety and Quality DiagnosticsBio-Rad Solutions

14-0686_FSD_Food_Safety_Booklet_Update_17933_FINAL.indd 1 4/10/14 11:13 AM

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


Food Safety and Quality Diagnostics • Bio-Rad Solutions | | 3


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


Tryptone-Salt/DiluentReady-to-use355-5754 9 ml x 25 tubes355-5756 90 ml x 6 bottles355-5796 3 L x 4 bags


30°C72 hrs

PCA Bacteria



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


Dilutions

TBX





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


RAPID’E.coli 2/AgarReady-to-use355-5297 200 ml x 6 bottles355-5299 100 ml x 6 bottles


TBX/AgarReady-to-use355-5309 100 ml x 6 bottles355-5307 200 ml x 6 bottles


Tryptose Lauryl Sulfate/BrothDehydrated356-4774 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




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


RAPID’E.coli 2

Dilutions

Tryptose LaurylSulfate

EC Medium

Peptone Water

Indole test reading and calculation


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


* 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




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



BGBLB (Brilliant Green BileLactose Broth)/BrothReady-to-use357-8024 10 ml x 25 tubes



RAPID’E.coli 2/AgarReady-to-use355-5297 200 ml x 6 bottles355-5299 100 ml x 6 bottles


Tryptose Lauryl Sulfate/BrothDehydrated356-4774 500 g



VRBL (Violet Red Bile Lactose)/AgarReady-to-use355-4799 100 ml x 6 bottles


VRBL Coliforms




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*


RAPID’E.coli 2

Enumeration of characteristic colonies

and confirmation for atypical colonies


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


(2 X)*

ColiformsAlternative method: RAPID’E.coli 2 Agar

RAPID’E.coli 2 E. coli Other Coliforms

Total Coliforms




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





EE/Broth Dehydrated356-4794 500 g

Nutrient (2.8%)/AgarReady-to-use355-4478 8 ml x 25 tubes


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




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%)


VRBG


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*


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



RAPID’Enterobacteriaceae/AgarReady-to-use355-4012 200 ml x 6 bottles





Pathogen Testing


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


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



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


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


andTretiationate

Broth

Modified Rappaport

Vassiliadis Brothand

TetrathionateBroth


Pathogen Testing


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


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


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


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




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


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


Salmonella Latex Kit

Detection of groups B to E and G355-6710 75 tests



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


Hektoen/Agar Ready-to-use355-4386 100 ml x 6 bottles356-3894 90 mm x 20 dishes


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


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


TSI (Triple Sugar Iron)/AgarDehydrated356-4384 500 g

XLD (Xylose Lysine Desoxycholate)/AgarReady-to-use354-1751 90 mm x 20 dishes


XLT 4 (Xylose Lysine Tergitol 4)/AgarReady-to-use356-3654 90 mm x 20 dishes

RAPID’Salmonella

Salmonella spp.

Other Enterobacteriaceae


Pathogen Testing






iQ-Check Cronobacter spp. 357-8137 96 tests

RAPID’Sakazakii/AgarReady-to-use356-3971 90 mm x 20 dishes


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



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


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.


Confirmation of positive with RAPID’Sakazakii

+/-

37°C18 hrs ± 2 hrs

Infant formula

37°C20 hrs ± 2 hrs

37°C4 hrs ± 1 hr


Pathogen Testing


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.



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


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



Fraser ½/BrothComplete Medium355-5797 225 ml x 6 bottles355-5794 3 L x 4 bags355-5792 5 L x 2 bags


Supplement356-4616 10 vials

Fraser 1/Broth Complete Medium355-4569 10 ml x 25 tubes



Oxford/AgarReady-to-use356-3664 90 mm x 20 dishes

Palcam/AgarDehydrated Base356-4754 500 g


EN ISO 11290-2/A1 (02/2005)

Horizontal method for the detection and enumeration of L. monocytogenes

Part 2: Enumeration method

Listeria spp.


Listeriamonocytogenes


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)


Pathogen Testing


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.


L.monocytogenes


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


35°C24 – 48 hrs

35°C24 – 48 hrs


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



Pathogen Testing

Listeria monocytogenes - Alternative method: RAPID’L.mono Agar

Detection of Listeria monocytogenesNF VALIDATION according to ISO 16140


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


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


Directenumeration


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


Pathogen Testing


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


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


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



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.




Rhamnose Test355-3669 28 tests




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


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



Pathogen Testing




Fraser ½/BrothReady-to-use355-5797 225 ml x 6 bottles355-5794 3 L x 4 bags355-5792 5 L x 2 bags



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 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


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.


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



Pathogen Testing





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


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



Pathogen Testing


Escherichia coli O157:H7 - Alternative method: iQ-Check E.coli O157:H7 real-time PCR Kit


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




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


41.5°C24-48 hrs

Innoculate a steril loop

mCDD Agar Other solid selective medium

Isolation Columbia blood Agar


Study of growth Columbia blood Agar


Oxidase

25°C44 ± 4 hrs


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




Oxidase

25°C44 ± 4 hrs


41.5°C44 ± 4 hrs

(aerobic atmosphere)

Examination of morphology and motility

Dilution

41.5°C40-48 hrs


Select 5 colonies


Pathogen Testing


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

+




Pathogen Testing

Campylobacter spp. - Alternative method: RAPID’Campylobacter spp.

Sample enrichment Suspension/Dilution





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


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


41.5 °C ± 1°C(24 hrs) 48 hrs


RAPID’Campylobacter Campylobacter spp.


Pathogen Testing


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


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



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


Enumeration and Confirmation

Sample

Confirmation

Pastorex™

Staph Plus

Spot on Baird-Parkerfor RPF

or



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


RPF Supplement356-4618 10 vials

BHI (Brain Heart Infusion)/BrothReady-to-use355-3664 10 ml x 25 tubes


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


Spread 1 ml on 3 platesBaird Parker

Dilutions

35°C45-48 hrs

RAPID’Staph

37°C24 hrs ± 2 hrs


Pathogen Testing


Clostridium perfringens

NF EN ISO 7937 (02/2005)

Horizontal method for the enumeration of


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


LactoseSulfite

MobilityNitrate

LactoseGelatine

Clostridiumperfringens(yellow/gas+)

Clostridiumperfringens(red coloration/gas+)

Clostridiumperfringens(black precipitate/

gas+)

37°C18-24 hrs

Anaerobescondition

TSC

TSC (add D-cycloserine)



Lactose Sulfite/BrothBase356-4914 100 g

Sheep Blood/AgarReady to use356-3784 90 mm x 20 dishes


Sheep Blood Supplement355-6652 50 ml vial

Thioglycolate + Resazurin/BrothReady to use356-3613 10 ml x 25 tubes


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



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


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%)


Biochemical and serologicalconfirmation



Hektoen/AgarReady-to-use355-4386 100 ml x 6 bottles356-3894 90 mm x 20 dishes


Mac Conkey + Crystal Violet/AgarReady-to-use356-3617 90 mm x 20 dishes



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


Mossel (MYP) Bacillus cereus


(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.


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



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


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




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


© Bio-Rad 10 17933 14-0686 Rev I 04/2014

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Food Safety and Quality Diagnostics - Bio-Rad · PDF fileFood Testing Food Safety and Quality Diagnostics Bio-Rad Solutions 14-0686_FSD_Food_Safety_Booklet_Update_17933_FINAL.indd