Applying USP Guidelines as Part of a Comprehensive RMM ValidationGoals of Validation Architecture Delivers meaningful and relevant content to end user Can e ectively determine that the performance characteristics of the RMM meet the requirements for the intended application, in comparison to the traditional method (USP)

Harmonizes statistical approach and methods that satisfy relatedguidelines (e.g. EP, ISO, TR33, JIS...)

Methods and results withstand the rigor, scope, and neutralityexpected by Regulators

Creating the right RMM validation approach begins with the righthuman resources and connections: Prospective users Industry thought leaders Research experts Microbiologists

Engineers Scientists Biostatisticians

People

USP is the benchmark guidance for RMM validation, butother relevant guidance documents should be considered andharmonized for a comprehensive validation:

Guidance &Regulations

Unique procedures for RMM testing are often integral to success, but they should build upon best practices, ensure traceability, and be formalized:Microbes (ATCC strain #)Bacillus atrophaeus (9372)Corynebacterium afermentans (51403)Escherichia coli (13706)Staphylococcus epidermidis (12228)Micrococcus lylae (27566)Aspergillus niger (9142)

Procedures & Microbes

RMM

ISO

13485

14644

21501

JIS 3836

9921

TR33

21CFR

EP

USP

Part 11

1223

5.1.6

Guidance &RegulationsPeople Statistics

Data &AnalysisEquipment

Rapid MicroMethodsValidation

USPEPISO

Procedures & Microbes Analysis

BioVigilants Comprehensive Approach

To e ectively apply a Rapid Microbial Method (RMM) technique and leverage its value, the end user must match his or her application and requirements to the bene ts and performance of the RMM.

Understanding the general approach to validation as well as the inter-relationships between personnel and their quali cations, guidance and regulations, statistics employed, test equipment, test protocols, and data analysis techniques enhances this evaluation. Through this holistic understanding, more robust dialogue can be fostered, and ultimately, a more successful adoption of an RMM technology may be realized.

USP: The Benchmark Guidance for RMM Validation

Scott Morris Applications Engineering ManagerBioVigilant Systems, Inc.

Not Speci ed in USP Universal minimum dataset requirements Tiered approach for acceptance criteria Evaluating/validating test environment/collection method Handling anomalous and nonquantitative data results Testing of multiple instruments for validation (reference and alternative instruments) Accommodating and/or circumventing the limitations of reference (traditional) methods

Prescribed Validation Criteria (and Utilized by BioVigilant)

Validation of an RMM is not something that merely can be claimed; it must be demonstrated.

Visit BioVigilants Booth 14 to obtain your copy of IMDA USP Test Results.

Testing of an RMM often requires specialized equipment and stateof theart facilities; these components should be calibrated andthoroughly characterized for a rigorous assessment. Multimillion dollar purposebuilt bio test facility and automatedaerosol test chamber

Please refer to PDA Poster entitled, Development and Validation of a Test Chamber by Shinsuke Yamasaki, Azbil Corporation

Equipment

IMD-A 300 (2)IMD-A 350 (2)SAS, SMA, MASKanomax LSAPC

Alternate Methods

Reference Methods

Test Chamber

The goal of the analysis and nal data should be to distill the value ofthe other ve validation components to clearly demonstrate whether equivalence or better was achieved and deliver meaningful and relevant data to the end user.

Data &Analysis

IMD-A 300 (#1) IMD-A 300 (#2) IMD-A 350 (#1) IMD-A 350 (#2) SAS MAS SMA

ConcentrationT1 T2 T3 T4 T5

Biolog

ic Cou

nts or CFU

Per Lite

r

1000.00

100.00

10.00

1.00

0.10

A comprehensive statistical architecture should be created to provide meaningful analysis, and also be detailed and formalized to ensure a repeatable and unbiased approach.

Statistics

General Themes and Methods for a Robust Statistical ArchitectureTiered Acceptance Criteria Using a primary criteria and alternate criteria for evaluation of statistical signi cance allows for a

simpli ed non-inferiority type of approachParallel Sample Collection All instruments should collect data simultaneously to minimize systematic error and allow pairing

of replicates in analysis (e.g. paired T-test)Uniform Dataset Conditioning Consistent rules applied to the conditioning of data and treatment of unsuitable (e.g. TNTC)

replicates ensures an unbiased and consistent approachMultiple Alternate and Reference Instruments

Using multiple instruments creates a more robust evaluation, ensures data has end-user relevance, and allows for straightforward assessment of Ruggedness

Minimum Number of Replicates

USP does not specify consistent minimum replicates (if at all), but a universal minimum allows monolithic testing and also enhances statistical signi cance of the dataset

Range of Quantitation De ning an ROQ not only provides guidelines for creating optimal test conditions, it provides a consistent method for ensuring an instrument is operating acceptably for data analysis

Appropriate Data Transforms Data transforms [e.g. log10(x)] are useful in statistical analysis, but their suitability should be evaluated and the assumption of data normality avoided

The graph to the left summarizes aerosol chamber results for a Bacillus atrophaeus microbe across ve concentrations. It clearly demonstrates not only the capability of the chamber to produce repeatable and homogeneous results, but it illustrates how the IMD-A systems yield repeatably higher recoveries than the reference methods

Bacillus atrophaeus

To help achieve these goals BioVigilant researched and employed the use of someunique statistical tools and methods: Winsorization Paired TTest Levene Test

Binomial Test Independent Linearity Reference Precision Transform

Alternative Acceptance Criteria

By the Numbers:> 4*108 Microbes aerosolized

2880 Individual samples collected

> 1100 Agar plates manually counted

1460 USP evaluation 34 Tears cried

Design of Experiment:9 USP Metrics

8 Instruments (4 x IMD-A)

6 Microbes5 Concentrations

12 Replicates