1. AOAC & Industry Perspectives: Validation and Verification of Alternative Microbiological Methods
Ronald Johnson
Senior Manager
Scientific Affairs - bioMerieux Industry
AOAC INTERNATIONAL Board of Director

2. Roles at bioMerieux and AOAC
bioMerieux (bMx) Project Management: AOAC Validations North America Scientific Liaison: Government/Industry AOAC AOAC International Board Director (BOD) Past Chair - AOAC Research Institute Board Director Past Chair - AOAC RI Advisory Council Study Director- bioMerieux method validations Method author: Journal of AOAC, Inside Laboratory Magazine

3. AOAC INTERNATIONAL
Non-profit organization with international membership
Consensus organization - Headquarters Gaithersburg, MD
Established in 1884 to review, validate &
publish analytical methods. Formerly part of FDA.
125th Anniversary in 2009!
best known in food and agriculture
chemistry and microbiology methods for Official Methods of
AnalysisSM status (OMA) “Big Book of Methods” & online
OMA methods are recognized in the U.S. Code of Federal
Regulations and are legally defensible in court worldwide
Subsidiary - AOAC Research Institute,
established 1991

4. AOAC is Staying Relevant
Methods validation: AOAC Perspective
AOAC is Staying Relevant
Globally recognized, non-profit Association
Over 3000 members from government, industry, and academia
15 Active AOAC Sections Worldwide

5. Harmonized Performance
Method Consensus with Global Stakeholders
Harmonized Performance
&
Acceptance Criteria
CFIA, HPB,
FDA BAM,
USDA MLG,
ISO
AFNOR
CEN
MicroVal
Food Manufacturers
AOAC OMA, AOAC RI
Test Kit
Manufacturers

6. AOAC INTERNATIONAL Current Projects
Stakeholder Panel on Infant Formula and Adult Nutritionals (SPIFAN)
Stakeholder Panel on Strategic Food Analytical Methods (SPSFAM)
International Stakeholder Panel on Alternative Methods (ISPAM)
ISO harmonization, Produce, STEC
DHS - Stakeholder Panel on Select Agent Detection Assays (SPADA)
Stakeholder Panel on Infant Formula and Adult Nutritionals (SPIFAN)
The global infant formula community is in the process of developing voluntary consensus standards for global dispute resolution methods. With consensus developed standards for the first set of nutrients, expert review panels will review and adopt methods as AOAC Official Methods of Analysis for these nutrients.
Stakeholder Panel on Strategic Food Analytical Methods (SPSFAM)
Food companies are coming together to lead the charge to develop the much needed standards and methods that promote food safety and facilitate trade. Initial areas of interest include antioxidants, flavanoids, contaminants and raw ingredients.
International Stakeholder Panel on Alternative Methods (ISPAM)
Food companies use analytical methods to meet regulatory and product safety testing requirements. The industry and government are seeking common ground for method validation, thereby promoting food safety and facilitating trade and yielding optimal efficiency. Developing harmonized internationally accepted method validation protocols for microbiology and qualitative chemistry methodology is the goal.
For information on SPIFAN, contact Anita Mishra, executive for scientific business development, at
For information on ISPAM, contact Krystyna McIver, senior director, stakeholder communications, at
For information on SPSFAM, contact Dawn Frazier, senior director, membership and education, at
For information on SPADA, contact Deborah McKenzie, senior director, standards development.

7. International Scope of Food Safety ISPAM: Harmonization
Microbiological methods are certified by official validation bodies to meet accepted performance standards set by official validation organizations.
The guidelines and criteria of AOAC in the U.S. and ISO in Europe are accepted in most of the rest of the world.
AOAC in the U.S. and AFNOR in Europe are the most widely recognized validation bodies.
AOAC International/ISO16140 Guidelines

8. Method Validation and Approval Organizations
AOAC PTM
AOAC PTM
AOAC OMA
AFNOR
Health
Canada
MicroVal
NMKL
NordVal
Acceptance of rapid method test kits across global
markets often depends on multiple validations

9. Regulatory Guidelines, Reference
Methods and Standards
AOAC PTM
Health
Canada
CFIA
USDA FSIS
AOAC Intl
FDA BAM
ISO
CEN
Global Companies may need to use different validations
with unique protocols at various production sites worldwide

10. The Weary Food Microbiologist
Health
Canada
CFIA
AOAC PTM
AOAC OMA
AOAC PTM
USDA FSIS
FDA BAM
Health
Canada
AFNOR
Help!!
AOAC Intl
ISO
MicroVal
NordVAl
CEN
So What is in your tool box of approved diagnostic test?
NMKL 10

11. Insanity?
Insanity: doing the same thing over and over again and expecting different results.”
Albert Einstein 11

12. BENEFITS OF METHOD HARMONIZATION
No Scientifically Valid Reason For Validating Food Microbiological Methods Differently
Need For Method Harmonization (ISO/AOAC) Has Been Recognized For Many Years with Limited Progress
Benefits of Harmonization:
- Broad Acceptance Of Microbiological
Methods And Associated Results
- Speeds Up Implementation Of New
Methods/Technologies Across Appropriate
Food Matrices
ISPAM Harmonization Efforts Ongoing

13. IMPACT OF GLOBAL TRADE ON THE WORLDWIDE FOOD SUPPLY
With increasing global trade there is an increasing concern about the safety and security of foods in the global supply chain 13

14. IMPACT OF GLOBAL TRADE ON FOOD SUPPLY
Consumer warnings:
Recalls and Outbreaks:
Company liability,
economic stability
Can often be prevented by
implementation of HACCP
and other food safety programs
and validating/verifying the
effectiveness of these programs
with AOAC Official (OMA)
methods/AOAC RI PTM methods 14

15. How to Better Safeguard
the Global Food Supply
“”Trust but Verify”
Verify method performance in your lab 15

16. Level of Confidence: Fit for purpose
Validated methods must be chosen according to the intended “fit for purpose for use” of the method.
AOAC validation requirements and lab verification should be appropriate for the intended “purpose for use” e.g.
Industrial process monitoring
Regulatory screening of foods
Industrial process verification
Regulatory compliance
Crisis management – outbreaks…traceback
Greater
Precision
required

17. Test Kit Manufacturer Perspective bioMerieux Goals for Food Safety
Provide Innovation to Food Industry
Shorter Turnaround Time is needed : Same Day solution
bioMerieux develop new technologies to reach this goal
Provide Performance to Industry
Sensivity & specificity enhancements
Phage detection
Provide Useful Tool to Industry
Integrated solution, with high throughput and limited hands-on-time
bioMerieux has a long history in automation of microbiology: VITEK2, VIDAS, TEMPO, BacT/Alert, AES

18. Test Kit Manufacturer Perspective Methods validation:
bioMerieux Development Process: Confidence in Performance Claims 1. Feasibility 2. Optimization Design Control Phases (controlled by FDA) 3. Internal Verification 4. External Verification 5. Industrialization 6. External Validation 7. Launch authorization 8. AOAC/AFNOR Validation

19. Test kit Co. Development & Validation Requirements for Pathogen Detection
1. Recognition of target analyte (Pure culture
Inclusivity/exclusivity)
Cell surface receptors (affinity/avidity) –
a. Antibody/antigen
b. Phage/conformational sites
c. Hybridization/PCR
Limit of detection of the target analyte
(spiked enrichment and detection)
growth dependent detection
a. 0 tolerance
b. 1 CFU/25g
c. 1 CFU/375g, 1 CFU/125g

20. AOAC (bMx) VALIDATION PROCESS
Inclusivity: Ability To Detect Target Analyte From A Wide Range Of Strains
-Minimum of 100 different serotypes for
Salmonella (>2200 serovars)
-Minimum of 50 for all other organisms
Exclusivity: Lack of Interference from a Relevant Range of Nontarget Strains
-Minimum of 30 isolates
Balance between sensitivity and specificity w/ near neighbors (priority: sensitivity)

21. Method Comparison (Core of Validation)
Evaluating a commercial kit in comparison to a reference method
Alternative method
Test sample
(Paired or unpaired)
Reference method

22. AOAC Official Methods (OMA)
VALIDATION PROCESS
Phase 1: Method Comparison (Pre-collaborative study) -Performed by one laboratory (contract lab) -All matrices claimed are evaluated -AOAC Performance Tested SM (PTM) -Independent laboratory Phase 2: Interlaboratory Collaborative Study Multiple laboratories (minimum of 10 labs)

23. AOAC Validation Programs
AOAC International
Official Methods of AnalysisSM (OMA)
12 months for approval
AOAC Research Institute
Performance Tested MethodsSM (PTM)
6 months for approved certificate
Harmonized Program
PTM + OMA (12 approval for both) + 23

24. bioMerieux AOAC Approvals
21 OMA approvals (all foods/variety of foods, env.)
14 VIDAS- rapid pathogen detection 2 TEMPO- automated QI testing 2 VITEK 2- rapid identification 1 API
2 VITEK (legacy) rapid identification 2

25. bioMerieux AOAC Approvals
27 AOAC RI – PTM approvals (variety of foods, env.)
11 VIDAS 6 TEMPO 3 VITEK
6 Adiafood- PCR
2 Plate methods ALOA, CFA 2

26. Food Safety Modernization Act (FSMA)
•Mandatory Food Facility Registration •Issuing recalls: Direct FDA recalls of food products. •Conducting inspections: more frequent inspections based on risk •Importing food: Greater oversight •Preventing problems: HACCP plan in place •Focusing on science and risk

27. As a microbiologist, as an analyst in food safety the job can be overwhelming, downright huge, but don’t feel small by getting lost in the data. David and goliath battle big brother

28. ISO Requirements for Verification
Importance of 3rd Party Validated Methods as it Relates to ISO Laboratory Accreditation

29. ISO Requirements for Verification
Official or reference (validated) methods already published and intended for specific matrices and target organisms that will be incorporated “as is” into the current method document format do not need to be revalidated. However, the ability to perform the analysis must be verified to ensure that the method is fit for its intended purpose

30. ISO Requirements for Verification
In Addition: Any modification to an official/reference method, including extension to a new matrix or target organism requires revalidation/verification (Note: The level and rigor of verification will depend on nature/extent of the modifications)

31. Laboratory demonstrates the capability of
achieving specified method performance
characteristics before implementation and on an on-going basis.
Provides data to illustrate that the method is fit-for (it’s)-purpose i.e. meets customer
requirements

32. How much confidence do you need in method performance?
AOAC Validation Programs
How much confidence do you need in method performance?
PresidentTruman - Missouri famous plaque on his desk
Answer: SHOW-ME Validated and Verified “Fit-for-Purpose” Methods

33. Laboratory 17025 Accreditation
A procedure by which an authoritative body such as A2LA gives formal recognition that a laboratory fulfills specified requirements and is competent to carry out specific tasks.
Ultimately the aspect of accreditation that sets it apart from other quality recognition schemes (like ISO 9000 registration) is that it emphasizes the demonstration of competence to perform a task.
IEC is International Electrotechnical Commission, the international standards and conformity assessment body for all fields of electrotechnology.
SCC refers to the Standards Council of Canada
CAEAL refers to the Canadian Association of Environmental Analytical Laboratories
A2LA is the acronym for the American Association for Laboratory Accreditation
ISO 17025:2005 is the current international accreditation standard considered relevant for testing laboratories.

34. Technical Elements of ISO 17025
Personnel-Training
Separation of incompatible activities
Test Methods Verification Including
Measurement Uncertainty
Calculations and Data Transfers
Equipment
Computers or Automated Equipment
Protection of Data
Measurement Traceability
Sampling
Handling of Test Materials
Quality Control
Reporting the Results

35. Guideline applied: procedure in place
Big Picture of assessment for ISO accreditation:
1. Quality system
a. All components in place
b. Documentation
c. Traceability

36. ISO Requirements
1. Verification 2. Trending for measurement of uncertainty Proficiency testing (annually) AOAC, API
programs - Verify in house (initial verification of method for your scope) 20 data points 4, 5 analysts – 5 samples each Stable Matrix/target (NFDM, peanut butter, instant mash potatoes 1-5 CFU/25g)

37. ISO 17025 Requirements-continued
- Measuring uncertainty daily (daily control trending)
-track daily performance (pure cultures: pathogens, APC, Coliforms, enterobacteriaceae)
-Proficiency testing - Approved PT supplier (AOAC, API)
In the method scope: Verifying a similar matrix RTE turkey validated, but verifying RTE ham
-spike (i.e., 5-10 reps 2-5 cfu/25g)
Caveat: Use outside of method scope may require re-validaton!

38. ISO 17025 Method verification
AOAC 2008 Document
AOAC Analytical Laboratory Accreditation Criteria Committee (ALACC)

39. NOTE: Regulatory Guidance

40. Automated (bMx) Systems Easier to Accredit
VIDAS®, TEMPO®, VITEK®2
For test kits
Methodology

41. Automated (bMx) Systems Easier to Accredit
Summary:
Avoids transcription errors
Easy access to data
Data management for trend analysis/HACCP
Complete traceability of results
Simplifies audits
Connected to the LIMS
Historical data on process/products
Reproducible results
Compatible with ISO 17025:2005

42. Test Kit Manufacturer Perspective Final Thought:
The development of new testing technology presents a considerable challenge but this is not the end of the story.
No wine before its time!
Extensive work to achieve validation and laboratory implementation/verification is necessary before the new method is accepted.

43. Final Food for thought for ISO training
As an microbiologist, as an analyst in food safety the job can be overwhelming, downright huge, but don’t feel small by getting lost in the data. Thank you for your attention!

44. Networking Opportunities
AOAC Final thought: Take Advantage of the power of the AOAC Association!
Networking Opportunities
AOAC Annual Meeting
Section Meetings
Technical Divisions
Communities
Standards
Performance Tested MethodsSM
Official Methods of AnalysisSM
Publications
The Journal of AOAC INTERNATIONAL
Inside Laboratory Management Member Magazine
Laboratory Services
Laboratory Auditing
Proficiency Testing

45. AOAC International Expostion September 7 – 10, 2014Boca Raton, Flo
SCIENTIFIC SESSIONS
CALL for POSTERS
AOAC Section meetings: China, Europe, India, Japan, Latin American-Caribbean, Lowlands, Taiwan, Thailand, Mid-Atlantic, Mid-Canada, Midwest, Pacific Northwest, Pacific Southwest, Southern, Southern California

46. For more info: www.aoac.org
From the voice of bMx,
Thank you and good evening Anaheim!
On behalf of the AOAC International BOD have a great meeting and no wine before its time