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