Safety of Laboratory Staff and Quality Assurance Programmes

1. Safety of Laboratory Staff and Quality Assurance Programmes Lecture Module 11

2. General introduction

3. Laboratory safety Two basic objectives: • To ensure safety of laboratory staff • To ensure that no hazardous materials leave laboratory These are both achieved by well controlled safe working practices

4. Background information • WHO Laboratory Biosafety Manual 3rd edition • In addition, staff should conform to their national and institutional legislation on regulations regarding safe working generally in laboratories

5. Specifics There are someparticularfeaturesconcerningsafety in cytogeneticslaboratoriesthat are worthhighlighting. • Risk of blood borne infections • Riskfrom use of ultra-violet light • Riskfrom use of specialisedchemicals

6. Infection risk Two broad principles: • Containment of materials - Universal precautions • Staff health preventative measures - Vaccinations

7. Optical risk Ultraviolet lamps may be used in: • Sterilizing interior of class 2 microbiological safety cabinets • Exposing slides during FPG staining procedure • UV fluorescence microscopes

8. Chemicals classically used in biological dosimetry WHO Laboratory Biosafety Manual, part VI Certain fine chemicals and pharmaceuticals are used routinely in procedures of biological dosimetry When present in cultures or used in staining procedures, they are mostly used in small volumes and in dilutions that generally present no health hazard They are, however, made up and stored in concentrated stock solutions Main reagents of concern and their internationally agreed risk phrases (R numbers) are listed here

9. Chemical risk : meanings of internationally agreed risk phrases (R numbers) R20 Harmful by inhalation R21 Harmful in contact with skin R22 Harmful if swallowed R23 Toxic by inhalation R24 Toxic in contact with skin R25 Toxic if swallowed R26 Very toxic by inhalation R27 Very toxic in contact with skin R28 Very toxic if swallowed R36 Irritating to eyes R37 Irritating to respiratory system R38 Irritating to skin R40 Possible risk of irreversible effects R41 Risk of serious damage to eyes R42 May cause sensitization by inhalation R43 May cause sensitization by skin contact R46 May cause heritable genetic damage R61 May cause harm to the unborn child R63 Possible risk of harm to the unborn child

10. Quality programmes and the ISO standards

11. Overview of the technical validations

12. 2002 INTERNATIONAL INTERCOMPARISON

13. Standards used IAEA, EPR-Biodosimetry, 2011: Cytogenetic dosimetry: applications in preparedness for and response to radiation emergencies IAEA, Safety Guide GS-G-3.2: The management system for technical services in radiation safety ISO/CEI 17025:2005General requirements for the competence of testing and calibration laboratories ISO 19238:2004Radiation protection- performance criteria for service laboratories performing biological dosimetry by cytogenetics ISO 21243:2008Radiation protection — Performance criteria for laboratories performing cytogenetic triage for assessment of mass casualties in radiological or nuclear emergencies — General principles and application to dicentric assay

14. Standard must cover these aspects • What you are trying to achieve • What you do • How do you make sure it is right • What do you do when it is not right • And, lastly, how do you check that system works

15. ISO Standard general approach Why a quality assurance approach ? To satisfy the requirements of professionalism To point out its qualification and its competence To develop the competence by inquiring and informing Quality assurance approach can be divided into two parts: First, internal mechanisms such as “ internal audit” or “total quality management”, which are processes of professional self-review and quality awareness, respectively. Second, certification (accreditation) which applies to processes more under the direct control of managers and operated by an external agency, in other words an external standards approach. How ISO can help to achieve the standardization aims ? Large consensus from scientists to industrials Participation on a voluntary basis To develop the competence by inquiring and informing

16. Biodosimetry ISO documents

17. Interaction steps in handling numbers of cases

18. General organisation Client A satisfied client Measure and continual system improvement Corrective and preventive actions / Internal audits Client satisfaction measurement Management process Quality objectives, Management review Technical process Resources management Personnel department/ Purchases Supplies equipments andreagents Quality management system Documents control / Qualityand technical records control

19. One review of the system every year Check of all the documents to ensure current accuracy Quality objective checks Each staff member has their own function The head of the lab formulates the objectives and defines staff functions One technician is in charge of the quality assurance system One scientist is in charge of the laboratory material One technician is in charge of purchases One scientist is in charge of documents management Management process Quality objectives, Management review

20. Establishment of a complaints book: Difficult to fill in regularly Direct benefit for the lab Audit: To evaluate the system periodically To progress improvements Conducted by external consultants to have an original point of view Measure and continual system improvement Corrective and preventive actions / Internal audits

21. A quality assurance manual + others Traceability Of products Of the sample Equipment maintenance According to the critical points of the technique: Microscopes The laboratory (safety, sterility…) Quality management system Documents control / Qualityand technical records control

22. General institutional processes Recruiting Purchases: for some products only one supplier Materials management Identification of key products Guaranteed supply of ready-to-use products Resources management Personnel department/ Purchases Supplies equipments andreagents

23. Lab must ensure that it can be contacted 24 / 7 All contacts from potential clients should be followed up Results can be conveyed to client electronically to ensure speed Written report then follows and is endorsed by medical doctor The client is invited to give feed-back to laboratory Client Satisfied client Technicalprocess

24. List of documents linked to case investigation When medical doctor contacts the lab Fill in contact details form Send instructions sheet for blood sampling and transport conditions Send questionnaire for explaining accident circumstances (contract) When sample(s) arrive(s) Record of details of sample and its coding Record of culture details Record reagents specifications e.g., batch numbers

25. Technical validations A satisfied Client client Reference curve validation by statistic approach TECHNICAL PROCESS AND MAJOR VALIDATION STEPS Images analysis system Reference curve Reference curve validation validation validation Comparison of automatic scoring by by statistic approach statistic approach / manual scoring ( mitotic index and dicentric frequency ) Mean Mean dose dose received received by by the the Dicentrics Dicentrics Cell culture number number body body Cytogenetic dose Cytogenetic dose (Gy) (Gy) Method validation • other aspects : by experimental approach different , • Intra comparison parameters were tested Inter comparison

26. Approach through audits Uncertainty arises from: Products Material Operator Image analysis system Number of dicentrics Number of cells scored Yield of dicentrics Dose +/- 95 % CI Dose effect curve Dose Number of cells scored Curve fitting Dicentrics Dicentrics number number dose dose

27. Step 1 : Metaphase finder Step 2 : Manual scoring Step 1 : Metaphase finder Step 2 : High magnification acquisition Step 3 : Screen scoring Images analysis system validation Comparison of automatic scoring / manual scoring ( mitotic index and dicentric frequency ) • Aim: scoring 500, 1000 or 2000 cells as accurately and fast as possible • 2 ways:

28. The metaphase finder validation Comparison of the number of mitoses found by the system and the “real” number of metaphases 10 % error accepted High magnification validation Evaluation of scorable mitoses on the system Comparison of dicentrics yield by the system vs manual 10 % error accepted Images analysis system validation Comparison of automatic scoring / manual scoring ( mitotic index and dicentric frequency )

29. Method validation by experiments (1) Identification of potential influencing factors Identification of tested interval (GUM) Uncertainty on sample manipulation, dilution …

30. Parameters to measure Method validation by experiments (2) • The mitotic index • The dicentric frequency and the associated dose

31. Example of study of factors assessed for their impact on yield of dicentrics

32. Parameters to test Experimental range :

33. Screening plan of Plackett et Burman

34. Results for mitotic index Parameters which affect the result

35. Results for dicentric frequency No significant parameter

36. Summary of mitotic and dicentric indices Some parameters affect number of mitoses: BrdU concentration, medium volume, the blood volume, duration of culture, incubator temperature No parameter affects dicentric frequency No operator effect was measured for both mitotic index and dicentric yield

37. One slide scored (a low dose first year, a high dose second year) No more than 20 % variation Staff annual scoring proficiency check

38. Final thought: convince staff to follow QA&QC programme The main difficulties : to convince people of importance of such project to change their way of working all staff members have to be involved in project the method validation is time consuming generation of many documents To have efficient system it is important to build it as light as possible The benefits are improvement in : technical process quality raising standing of service in eyes of requestors and any legal outcomes Whole process is constantly evaluated and changes are made for improved efficiency