AOAC & Industry Perspectives:

Validation and Verification of Alternative Microbiological Methods

Ronald Johnson

Senior ManagerScientific Affairs - bioMerieux IndustryAOAC INTERNATIONAL Board of Director

bioMerieux (bMx)Project Management: AOAC ValidationsNorth America Scientific Liaison: Government/Industry

AOACAOAC International Board Director (BOD)Past Chair - AOAC Research Institute Board Director Past Chair - AOAC RI Advisory CouncilStudy Director- bioMerieux method validationsMethod author: Journal of AOAC, Inside Laboratory Magazine

Roles at bioMerieux and AOAC

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

AOAC is Staying Relevant

Methods validation:AOAC Perspective

Harmonized Performance

&Acceptance

Criteria

CFIA, HPB, FDA BAM,

USDA MLG,

ISOAFNOR

CENMicroValAOAC OMA,

AOAC RI

Food Manufacturers

Test Kit

Manufacturers

Method Consensus with Global Stakeholders

AOAC INTERNATIONAL Current Projects

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

Method Validation and Approval Organizations

AOAC OMA AFNORAOAC PTMAOAC

PTM

MicroVal

NMKL

HealthCanada

NordVal

Acceptance of rapid method test kits across global Acceptance of rapid method test kits across global

markets often depends onmarkets often depends on multiple validations multiple validations

Regulatory Guidelines, Reference Methods and Standards

USDA FSIS

AOAC PTM

CFIA

FDA BAM

CEN

AOAC Intl

HealthCanada

ISO

Global Companies may need to use different validations Global Companies may need to use different validations

with unique protocols at various production sites with unique protocols at various production sites worldwideworldwide

AOAC PTM

CFIA HealthCanada

FDA BAM USDA FSIS

AOAC Intl

CEN

ISO

AOAC OMA AOAC PTM

HealthCanada

AFNOR

MicroVal NordVAl

NMKL

Help!!

The Weary Food Microbiologist

Insanity: doing the same thing over and over again and expecting different results.”

Albert Einstein

Insanity?

•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

BENEFITS OF METHOD HARMONIZATION

With increasing global trade there is an increasing concern about the safety and security of foods in the global supply chain

IMPACT OF GLOBAL TRADE ON THE WORLDWIDE FOOD SUPPLYIMPACT OF GLOBAL TRADE ON THE WORLDWIDE FOOD SUPPLY

IMPACT OF GLOBAL TRADE ON FOOD SUPPLYIMPACT OF GLOBAL TRADE ON FOOD SUPPLY

Recalls and Outbreaks:

Consumer warnings:

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

“”Trust but Verify”

How to Better Safeguard the Global Food Supply

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 foodsIndustrial process verificationRegulatory complianceCrisis management – outbreaks…traceback

GreaterPrecisionrequired

Test Kit Manufacturer Perspective

bioMerieux Goals for Food Safety

Provide Innovation to Food IndustryShorter Turnaround Time is needed : Same Day

solutionbioMerieux develop new technologies to reach

this goalProvide Performance to Industry

Sensivity & specificity enhancements Phage detection

Provide Useful Tool to IndustryIntegrated solution, with high throughput and

limited hands-on-time bioMerieux has a long history in automation of

microbiology: VITEK2, VIDAS, TEMPO, BacT/Alert, AES

bioMerieux Development Process:Confidence in Performance Claims

1. Feasibility2. OptimizationDesign Control Phases (controlled by FDA)3. Internal Verification4. External Verification5. Industrialization6. External Validation7. Launch authorization8. AOAC/AFNOR Validation

Test Kit Manufacturer PerspectiveMethods validation:

1. Recognition of target analyte (Pure culture Inclusivity/exclusivity) Cell surface receptors (affinity/avidity) – a. Antibody/antigen b. Phage/conformational sites c. Hybridization/PCR

2. 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

Test kit Co. Development & Validation Requirements for Pathogen Detection

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)

AOAC (bMx) VALIDATION PROCESS

Reference method

Test sample

(Paired or unpaired)

Alternative method

Method Comparison (Core of Validation)

Evaluating a commercial kit in comparison to a reference method

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)

AOAC Official Methods (OMA) VALIDATION PROCESS

AOAC InternationalOfficial Methods of AnalysisSM (OMA)

12 months for approval-------------------------------------------------------------------------------------------------------------------

AOAC Research InstitutePerformance Tested MethodsSM (PTM)

6 months for approved certificate----------------------------------------------------------------------------------------------------------------------------

Harmonized Program PTM + OMA (12 approval for both)

+

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

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

•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

Food Safety Modernization Act (FSMA)

Importance of 3rd Party Validated Methods as itRelates to ISO 17025 Laboratory Accreditation

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 mustbe verified to ensure that the method is fit for itsintended purpose

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 willdepend on nature/extent of the modifications)

ISO Requirements for Verification

Laboratory demonstrates the capability ofachieving specified method performancecharacteristics before implementation and on an on-going basis.

Provides data to illustrate that the method is fit-for (it’s)-purpose i.e. meets customerrequirements

How much confidence do you need in method performance?

AOAC Validation Programs

Answer: SHOW-ME Validated and Verified “Fit-for-Purpose” Methods

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.

Laboratory 17025 Accreditation

Technical Elements of ISO 17025

• Personnel-Training• Separation of incompatible activities• Test Methods Verification Including

Measurement UncertaintyCalculations and Data Transfers

• EquipmentComputers or Automated EquipmentProtection of Data

• Measurement Traceability• Sampling• Handling of Test Materials• Quality Control• Reporting the Results

Big Picture of assessment for ISO accreditation:

1. Quality system a. All components in place b. Documentation c. Traceability

Guideline applied: procedure in place

1. Verification2. Trending for measurement of uncertainty3. 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)

ISO 17025 Requirements

- 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!

ISO 17025 Requirements-continued

AOAC 2008 Document

ISO 17025 Method verification

AOAC Analytical Laboratory Accreditation Criteria Committee (ALACC)

http://www.fda.gov/ScienceResearch/FieldScience/LaboratoryManual/ucm171877.htm#1

http://www.fsis.usda.gov/wps/wcm/connect/966638c7-1931-471f-a79e-4155ce461d65/Validation_Studies_Pathogen_Detection_Methods.pdf?MOD=AJPERES

http://www.fsis.usda.gov/wps/wcm/connect/d3382b4f-27df-4e69-9744-ea8d73a2d603/At-Least-Equal-to-Guide-MPI-Programs-Lab-Methods.pdf?MOD=AJPERES

NOTE: Regulatory Guidance

Automated (bMx) Systems Easier to Accredit

VIDAS®, TEMPO®, VITEK®2

– For test kits

– Methodology

Automated (bMx) Systems Easier to Accredit

Summary:

Avoids transcription errorsEasy access to dataData management for trend analysis/HACCP Complete traceability of resultsSimplifies auditsConnected to the LIMSHistorical data on process/productsReproducible resultsCompatible with ISO 17025:2005

Test Kit Manufacturer PerspectiveFinal 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.

Final Food for thought for ISO training

AOAC Final thought:Take Advantage of the power of

the AOAC Association!

AOAC International ExpostionSeptember 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

For more info:www.aoac.org

From the voice of bMx,

Thank you and good evening Anaheim!