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Validation and Verification of FlowcytometryDOUAA MOHAMMED SAYEDDIRECTOR OF FLOW CYTOMETRY LABSOUTH EGYPT CANCER INSTITUTE – ASSIUT UNIVERSITYDOUAA_SAYED2HOTMAIL.COM
1st Arab Flow cytometry working group workshop 14 th – 15th October 2016
Verification and ValidationWikipedia definition
Verification and Validation, in engineering or quality management systems, it is the act of reviewing, inspecting or testing, in order to establish and document that a product, service or system meets regulatory or technical standards.
Validation – “Are you building the right thing?”
Verification – “Are you building it right?”
Quality Assurance in FCM Why should we care?
Assurance that flow cytometry results you are generating are due to the properties of the sample and not changes in:
•Instrument •Assay performance
•Reagents •Operator variation
1st Arab Flow cytometry working group workshop 14 th – 15th October 2016
INSTRUMENT Basic Knowledge about the configuration of the machine is essential
It is important to know basic start up and shut down procedures along with routine maintenance processes
MAINTENANCE log has to be maintained
1st Arab Flow cytometry working group workshop 14 th – 15th October 2016
INSTRUMENT What all is to be monitored
◦ Optics ◦ Fluidics ◦ Electronics
How do you monitor◦ Manual methods◦ Automated methods
QC beads are very useful for monitoring and evaluating instrument performance
1st Arab Flow cytometry working group workshop 14 th – 15th October 2016
Flow Cytometry Beads Spherotech Beads
Cytometer Setup and Tracking (CS&T) beads
Seven colour setup beads
Calibrite Beads
Flow-Check™ Fluorospheres
IMMUNO-BRITE™ Standards Kit
Flow-Check™ Pro Fluorospheres
Cell Counting Beads
Antibody Capture beads
1st Arab Flow cytometry working group workshop 14 th – 15th October 2016
Key Performance Factors in High Quality Flow Cytometry Data
Relative measured values of fluorescenceLinearity and accuracy
Resolution of subpopulations, including dim subpopulationsSensitivity
Reproducibility of results and cytometer performanceTracking
Comparison of results across time and among laboratoriesStandardization
1st Arab Flow cytometry working group workshop 14 th – 15th October 2016
Flow Cytometry Assay
1st Arab Flow cytometry working group workshop 14 th – 15th October 2016
Flowcytometry Assay Verification
Confirm specifications established by the manufacturer
Accuracy
Precision
Reportable range of results
Verify of manufacturer’s normal range
ValidationIn-house, home brewed assay
• Accuracy• Precision• Analytical sensitivity• Analytical specificity• Reportable range• Reference range• Calibration procedures• Control procedures• Sample stability
1st Arab Flow cytometry working group workshop 14 th – 15th October 2016
Verification and Validation Lot to Lot Antibody VERIFICATION
Setting up a new permeabilisation and fixing procedure using home brewed reagents - Assay validation
1st Arab Flow cytometry working group workshop 14 th – 15th October 2016
1st Arab Flow cytometry working group workshop 14 th – 15th October 2016
80
ISO 15189The
international standard for medicallaboratory
quality and competence
Seeking for accreditation of Flow cytometry laboratory1st Arab Flow cytometry working group workshop 14 th – 15th October 2016
The ISO 15189 standard, designed specifically for medical laboratories, covers 15 management requirements and 8 technical requirements. The standard is concise and to the point, with each section consisting of anything from a few sentences up to several pages of explanation.
Management requirements Technical requirements
Organization and management Quality management system Document control Review of contracts Examination by referral laboratories External services and supplies Advisory services Resolution of complaints Identification and control of
nonconformities Corrective action Preventive action Continual improvement Quality and technical records Internal audits Management review
Personnel Accommodation and environmental
conditions Laboratory equipment Pre-examination procedures Examination procedures Assuring quality of examination procedures Post-examination procedures Reporting of results
1st Arab Flow cytometry working group workshop 14 th – 15th October 2016
First Accredited Flow Cytometry Lab Location: South Egypt Cancer Institute – Assiut Universityhttp://www.seci.info/flowcytometry/
1st Arab Flow cytometry working group workshop 14 th – 15th October 2016
1st Arab Flow cytometry working group workshop 14 th – 15th October 2016
Acknowledgements
Thank you1st Arab Flow cytometry working group workshop 14 th – 15th October 2016
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