IHC and process validation - NordiQC...
Transcript of IHC and process validation - NordiQC...
2nd NordiQC Conference on Applied Immunohistochemistry
Content
IHC and process validation
an aid towards standardization ?
▪ Background
▪ Definitions
▪ Process validation model
▪ Practical example
Content
▪ Background ▪ Definitions
▪ Process validation model
▪ Practical example
Background : lab challenges today
▪ Higher volume vs lower staffing ▪ More direct link to patient treatment ▪ Less patient material available ▪ Faster turn arround times (TAT) ▪ Lab specific settings vs standardization ▪ Quality management ▪ Higher demands from inspecting agencies
Background : “The Belgian situation”
Fed. Dept. of Health
▪ = official recognition of all pathology lab’s in Belgium
▪ Mandatory for every lab
▪ Royal Decree : 2011
▪ Practice guideline : 2014
Fed. Dept. of Economics
▪ ISO accreditation
▪ BELAC
▪ Not mandatory
▪ Specific tests within a scope
▪ ISO 15189
National accreditation programs moving to ISO 15189
Background : Practice guideline – ISO 15189
Technical Requirements Personnel
Accommodation and environmental conditions
Laboratory equipment, reagents, and consumables
Pre-examination processes
Examination processes
Post-examination processes
Reporting & release of results
Laboratory information management
Pre-analytical
Analytical Post - analytical
Management requirements Men Machine Means Methods Men Machine MeansMachi th dtM M
Laboratory Information System
Practice guide – ISO 15189
IHC : multistep & complex process
IHC Staining
Pre-analytical
Analytical Post - analytical
Pre-analytical Analytical Post -
analytical
Laboratoy Information System
Men Machine Means Methods
IHC Staining Tissue Fixation Preparation Sectioning Pre-treatment
Interpretation Control Cut-off level Reporting
Primary Ab Visualisation Staining platform
Process model Similar approach
IHC : from staining method to total process ?
IHC Staining
Pre-analytical
Analytical Post -
analytical Laboratoy Information System
Men : labtech’s, pathologists,…
Machine : processing, cutting, staining,…
Means : reagents, consumables, storage,…
Methods : fixation, processing, staining,…
IHC Staining Tissue Fixation Preparation Sectioning Pre-treatment
Interpretation Control Cut-off level Reporting
Primary Ab Visualisation Staining platform
M l bt h’ th l i t
Available info ?
▪ Validation of instruments
▪ Optimalization of methods
▪ Validation of methods
▪ IQC & EQC
▪ Reporting
▪ Etc …
▪ What about critical steps and/or influencing factors ?
▪ Known ? ▪ Effect ? ▪ Control ? ▪ Standardization ? ▪ Validation ?
▪ Total process ?
▪ Added value ?
IHC : from staining method to total process ? IHC : multistep & complex process - Questions
▪ Do we have confidence in our IHC process ?
▪ Is it capable of consistently delivering quality results ?
▪ Can we demonstrate that the process works as intended ?
▪ Can we detect a process drift ?
▪ Do we know the critical attributes throughout the process ?
▪ How many steps / influencing factors are controlled ?
IHC : multistep & complex process - Questions
▪ Confidence in process Failure rate ?
▪ Consistent process Standardization
▪ Process works as intended Meets acceptance criteria
▪ Critical attributes known What are the risk factors ?
▪ Influencing factors identified
Prospectiveveve ee risk analysis = == processss ss validationon
Process validation : new ?
▪ Process validation = Prospective risk analysis
▪ Used in pharma (GxP), clinical biology, industry, …
▪ Risk management integrated in quality management (Practice guidelines & ISO 15189)
▪ Different methods available
▪ Uses available information
▪ “Lab” tailored
Drivers for process validation
▪ Reduction of error through risk management and continual improvement
▪ Reduction of risk levels through risk reducing measures
▪ Higher degree of confidence in process
▪ Higher degree of standardization
▪ Higher degree of quality
Process validation = Framework for “in house” standardization
Content
▪ Background
▪ Definitions
▪ Process validation model
▪ Practical example
“Establishing documented evidence which provides a high degree of assurance that a specific process will consistently produce a product meeting it’s pre-determined specifications and quality attributes”
(FDA)
Definitions : VALIDATION
“Establishing documented evidence which provides a high degree of assurance that a specific process will consistently produce a product meeting it’s pre-determined specifications and quality attributes”
documented evidence
high degree of assurance
pre-determined specifications and quality attributes
▪ documentation
▪ confidence in the process
▪ validation criteria
Definitions : VALIDATION
“A series of actions or steps taken in order to achieve a particular end.”
▪Proces : – Manufacturer – User
▪ Integration / process design
Definitions : PROCESS
“Process validation is defined as the collection and evaluation of data, which establishes (scientific) evidence that a process is capable of consistently delivering quality products.”
Definitions : PROCESS VALIDATION
Definitions : PROCESS VALIDATION
Type of process validation Principle
Prospective/progressive risk analysis on total process
Concurrent during normal “production”, 3 batches monitored
Retrospective after process, using controls
Revalidation change in process
Types of process validation :
Prospective risk analysis
“Risk is the likelihood that there is damage, taking into account the extent of the damage.” - or - “Risk is the combination of the probability of occurrence of harm and the severity of that harm.”
(ISO/IEC Guide 51).
(≠ Hazard = “a potential source of harm”)
(≠ Hazard = “a potential source of harm”)
Definitions : RISK
Content
▪ Background
▪ Definitions
▪ Process validation model ▪ Practical example
Process validation method : steps
▪ Process validation = prospective risk analysis
▪ Steps : 1. Define scope for process validation 2. Describe process 3. Determine risk criteria 4. Perform risk assessment 5. Determine validation criteria 6. Validate appropriate risk factors/acceptance criteria 7. Maintain in a validated state
1. Define the scope for validation
IHC Staining
Pre-analytical
Analytical Post -
analytical
Interpretation Control Cut-off level Reporting
Tissue Fixation Preparation Sectioning Pre-treatment
Primary Ab Visualisation Staining platform
IHC Staining SAMPLE – REQUEST FOR STAINING
REPORT
1. Define the scope for validation
IHC Staining
Pre-analytical
Analytical Post -
analytical
Interpretation Control Cut-off level Reporting
Tissue Fixation Preparation Sectioning Pre-treatment
Primary Ab Visualisation Staining platform
IHC Staining process
Confirmation testing Correlation IHC - ISH
• Reagent storage • Reagent acceptance testing
• Instrument maintenance • Instrument acceptance
testing
• Time to fixation • Transport monitoring
• Processing
• Control preparation • Control storage
• Histotech - pathologist • Labmanagement • Quality management
• Lab. Information System
SAMPLE – REQUEST FOR TESTING
REPORT & FOLLOW UP OF RESULTS
1. Define the scope for validation
IHC - Single stain/panel IHC - Double stain
Kappa/ Lambda doublestain BioCare Medical K/L prim. Ab cocktail + Leica ChromoPlex™ 1 Dual Detection
K
L
CD3 – CD20 – Bcl 2 Leica Bond RTU + Leica Bond Refine Detection
CD20 Bcl2
CD3
1. Define the scope for validation
IHC + confirmation testing ISH - SISH
HER-2 (4B5) Roche RTU Confirm HER-2 + ultraView Universal DAB detection
HER-2 – Silver In situ Hybridization Roche Dual ISH HER-2 + SISH DNP Silver & Red DIG detection
1. Define the scope for validation
PCR IHC + confirmation testing
MSH 6 (clone 44) Becton Dickinson 1/400 + Leica Bond Refine Detection
2. Describe process for validation
▪ Process description : ▪ Main process ▪ Sub process ▪ Combination
▪ Method : ▪ Text ▪ Scheme : Word™, Visio™,…
= systematic overview process within scope
2. Describe process for validation
Sample
Histochemical Immuno Molecular
Histology Cytology
Procesflow !
2. Describe process for validation
Procesflow !
RequestLIS
Selectsamples
Printtasklist
LIS
Printlabel
barcode
Sample oncoated slide
Controlslideand/orsample
PerformRun
YES
NO
Store slide<24 – 48u :RT> 48u : -20°C
Load slides Load reagents
Start run
Scanslides andreagents
Performstaining
Bluing &mountingEnd of staining
Correctreagents
Pathologist Reporting
Removereagents
GrossingSamplereceptionSample
Microscopy
Processing &embedding Sectioning
3. Define risk criteria
▪ General requirements : – Legal – Standards : e.g. ISO 15189
▪ Practical experience, GLP
▪ Guidelines : e.g. ASCO/CAP, CLSI, …
▪ Literature
▪ Recommendations : – EQC organisations e.g. NORDIQC – manufacturers
• Manufacturer • Acceptance
testing
• Optimalization
• Validation • Maintenance • Reagent
management
• Training
Men Machine
Means Method
Input ? Depends on scope !
3. Define risk criteria
Available info
▪ ASCO/CAP HER-2 guidelines : – Fixation time 6-72 h
Used info
▪ Fixation time 6-72h
dd fi ff
3. Define risk criteria : possible risks
▪ Determine possible sources of errors - risks ▪ What can go wrong ? ▪ Not complying to guidelines ?
▪ Risk identification : input ? ▪ Brainstorming ▪ Fault tree analysis ▪ Fishbone diagram ▪ …
3. Define risk criteria : possible risks
Fish bone diagram (Ishikawa)
Scope
Men
Method
Machine
Means
Primary
Secundary
Scope : cause & effect • Brainstorm • using fishboneand “M”
model 1st line risk 2nd line related risk
4. Perform risk assessment : FMEA
▪ FMEA : Failure Mode and Effects Analysis
▪ Risk = chance x effect values ? scoring ?
Processtep Failure Consequence Describe processtep What can go wrong ? What are the consequences ?
Grossing : • Time to fixation
Time to fixation > 1h
Impact on HER-2 staining
Processflow Fishbone diagram INPUT
▪ likelihood
▪ Probability
▪ Frequency
▪ likelihoood Common Terms Rating Example (ISO 14971)
Practical Example
Frequent 5 ≥ 1/1,000 > 1x/week
Probable 4 < 1/1,000 and ≥1/10,000
Once every few months
Occasional 3 < 1/10,000 and ≥1/100,000
Once a year
Remote 2 < 1/100,000 and ≥1/1,000,000
Once every few years
Improbable 1 < 1/1,000,000 and ≥10,000,000
Unlikely to ever happen
4. Perform risk assessment : scoring criteria
Rating : • 1-5 • 1-6 • 1-10 • …
▪ extent
▪ severity
Common Terms Rating Possible Description (ISO 14971)
Catastrophic 5 Results in patient death Critical 4 Results in permanent injury of life-threatening
injury Serious 3 Results in injury or impairment requiring
professional medical intervention
Minor 2 Results in temporary injury or impairment not requiring professional medical intervention
Negligible 1 Inconvenience or temporary discomfort
4. Perform risk assessment : scoring criteria
Rating : • 1-5 • 1-6 • 1-10 • …
Risk = chance x effect
+ Detectability
= Risk Priority Number
Priority & acceptance levels
Common Terms
Rating Example
Low 5 Control is ineffective 4 Control less likely to detect the
failure 3 Control may or may not detect the
failure 2 Control almost always detects the
failure High 1 Control can detect the failure
4. Perform risk assessment : scoring criteria
Patient safety
• Sample mismatch • Staining mismatch • Interpretation of results
Integrity of data
• Case, task : number • Data transfer
Continuum of services
• Maintenance • Stock management • …
4. Perform risk assessment : scoring criteria
▪ Proces description (processflow)
▪ Risk criteria – input - scope (fishbone diagram)
▪ Risk / failure : patient safety, data integrity, continuity of services
Processtep Failure Consequence Chance Effect Risk Detection RPN
Describe processtep
What can go wrong ? What are the
consequences ? (1 – 6) (1 – 6)
(1 – 6)
Grossing : • Time to
fixation
Time to fixation > 1h
Impact on HER-
2 staining 2 6 12 2 24
4. Perform risk assessment : FMEA scoring
5. Select validation criteria
RPN (Risk Priority Number) : (= Risk x detection)
• Allows ranking
• Allows selection of validation items
• Level of detection possibilities : • Visual, manual • Automated
5. Select validation criteria
RPN (Risk Priority Number)
Unacceptable 60 Class 1 High 21 - 60 Class 2 Medium 11 - 20 Class 3 Low 1 - 10 Class 4
RPN Class Further action required ? Acceptance ? Class 1 Direct action No Class 2 Short term action No Class 3 Middle to lang term action Yes, within context Klasse 4 None Ja
Validation criteria !
Determine acceptance level !
6. Perform validation
▪ FMEA : Selected risks - RPN = validation
▪ Use available information, including : – Validation of instruments – Validation of methods – Keeping instrments & methods in a validated state
▪ Perform validation on “new” items
▪ Determine acceptance criteria validation
7. Maintain in a validated state = active !
▪ Instruments : – Validation – Maintenance – Control T°
▪ Methods : – Validation – Controls
▪ Proces validation monitoring
▪ Change management
Content
▪ Background
▪ Definitions
▪ Process validation model
▪ Practical example
Scope = HER-2 testing
RequestLIS
Selectsamples
Printtasklist
LIS
Printlabel
barcode
Sample oncoated slide
Controlslideand/orsample
PerformRun
YES
NO
Store slide<24 – 48u :RT> 48u : -20°C
Load slides Load reagents
Start run
Scanslides andreagents
Performstaining
Bluing &mountingEnd of staining
Correctreagents
Pathologist Reporting
Removereagents
GrossingSamplereceptionSample
Microscopy
Processing &embedding Sectioning
▪ Input - plug in :
▪ Guidelines : Belgian guidelines on HER-2 testing, ASCO/CAP
▪ Literature references
▪ Guidelines from manufacturer
▪ Good laboratory practice
▪ Validation of instruments & methods
HER-2 testing : risk criteria HER-2 testing : risk criteria
HER-2 testing : FMEA risk assessment
▪ FMEA :
Code Process step Failure Consequence Chance Effect Risk grade Detection RPN
Score Number % Follow up ?
Unacceptable 1 1,67% Yes
High 13 21,67% Yes
Medium 8 13,33% No
Low 38 63,33% No
Total 60 100%
HER-2 testing : FMEA : results
▪ FMEA : 14 validation points : – 1 unacceptable – 13 high risk
▪ = validation items
▪ Risk reducing measures incorporated in validation
HER-2 testing : validation criteria
▪ Validation of instruments
▪ Validation of methods
▪ Validation of LIMS
▪ Internal quality control
▪ Acceptance testing critical reagents
▪ Monitoring critical temperatures : – Stretching tables – Paraffin stations – Refrigirator & freezer
Allready validated ! ▪ Maintaining the validated state of instruments
▪ Maintaining validated state of methods
▪ Appropriate use of appropriate controls
▪ Monitoring critical process point
HER-2 testing : Maintaining a validated state
▪ Process validation will - in a systematic and documented way - highlight critical points within the scope of the IHC process. The validation and follow up of these points will, toghether with the validated IHC method, be an aid to the standardization of the total IHC process.
Conclusion
THANK YOU !