1. Quality Management -Standards
Joan Kelley
28th Nov 2002

2. Background Employed UK MoD Assessor in QC system
Transferred to UKAS (then NATLAS) on inception
Testing lab UKAS (now ISO 17025) Accredited since 1984

3. Accreditation Bodies UKAS-UK Accreditation Service
NAB-National Accreditation Board,Ireland
BELCERT and BELTEST, Belgium
Comite Francais d’Accreditation
Deutscher Akkreditierungsrat
Raad voor Accreditatie

4. Cooperative Bodies European co-operation for Accreditation (EA)
EA 4/10 Accreditation for Microbiological Laboratories, July 2002
accreditation.org
International Laboratory Accreditation Cooperation (ILAC)

5. Context
Regulations = EN45001, European Standard (others GLP, ISO 9000 etc)
But since early 2000………….
ISO/IEC came into force
General Requirements for the Competence of Testing and Calibration Laboratories

6. Current Standards ISO 17025- Testing and ‘methods’
ISO 9000 series-production and management systems
National and international GLP standards-Medical research

7. Change in Philosophy Process driven approach
-does the system work, rather than -does it contain specific features
More flexibility for the lab.

8. Format Most technical standards have two main sections ……………...
Management requirements
Technical requirements

9. Management
If part of larger company, key personnel for lab must be defined
Lab must have tech management but can be a team-individual not necessary

10. Documents Quality Manual Methods (Technical Operating Procedures)
Standard Operating Procedures

11. Quality System Management review Audit programme Document control
Control of complaints and anomolies
Clear job descriptions
Training records
etc etc etc

12. Contract Review Lab must have procedures which ensure that……
methods to be used are defined,documented and understood
lab assured it has capability and resources to carry out work
appropriate methods are used to meet clients needs

13. Preventive Actions
Pro-active look for opportunities to improve not just corrective actions
Can come from……..
Internal audits,review of AQC data,staff suggestions,management review

14. Technical requirements ISO 17025
Increased emphasis on uncertainty of measurement, always a minefield for microbiol.
In house calibrations given emphasis
Future training to be addressed
Opinions and interpretations can be accredited

15. Application of Standards to Microbiology labs
Some Guidelines

16. Quality Control
Internal AQC to monitor individual performances,cross checks between microbiologists,duplicate preps. of single samples by different people.
QC of batches of media and diluents ( more later)

17. Inter-laboratory comparisons
Lab should participate in external AQC schemes when available and appropriate-Collections to establish system??
eg PHLS Water and Food QC, FAPAS,NEQAS, SMART etc etc
Must be seen to act on results-don’t just file and forget

18. Staff Senior member of staff must have appropriate qualifications
Alternatives may be acceptable with extensive experience

19. Equipment Records for all essential equipment
Cleaning programmes for appropriate equipment together with documented records
eg water baths, incubators, fridges, freezers

20. Reagents Generally ANALAR Water -deionised or distilled
Regents labelled, identity, opening and expiry dates,conc, storage requirements
Includes all media and diluents

21. Calibration
Must have a co-ordinated, recorded programme encompassing all lab activities
Measurements of significance to methods must be traceable to national/international standards

22. Temperature Measuring Devices
Fridges, incubators,water baths,drying ovens, autoclaves
Where temp needs to be better than +/- 1C calibrated thermometer in appropriate range essential
Otherwise BS 593 may be adequate or calibrated ref used to check working thermometers

23. Cont……. Ref thermometers, calibrated every 5 years
Annual ice point check
Thermocouples-traceable to national standards
Working thermocouples can be calibrated in house against ref standards

24. Weights and Balances
Must be calibrated at least annually depending on use and environment
In house or by Accredited service
Daily drift checks

25. Volume
Pipettes etc must be calibrated and traceable to national standards
Variable volume automatic pipettes discouraged
Disposable pipettes should be batch checked even if supplied from ISO manufacturer

26. pH Checked daily or before use Buffers from quality supplier
Buffers prepared to standard and stored appropriately with an expiry date

27. Humidity
Must be calibrated and traceable
Easiest to send to experts!

28. Autoclaves
On purchase-multi point validation for all typical loads and put results to use!
Traceable to national standards
Thereafter calibration of timer and temperature measuring equipment annually
Again act on the results

29. Cont Records of all loads and runs must be taken and kept
In addition load monitors must be used with each load

30. Air velocity
If used in eg safety cabinets (ie not done under contract) must be checked for accuracy at regular intervals.

31. Method Validation
Methods developed in house, modifications and applications of standard methods must be validated
But also need to prove you can get the correct results even in standard tests

32. Essential Features Limits of detection Selectivity Precision
Reproducibility
Procedures depend on requirements/qualitative or quantitative/new or existing method?
Reference materials

33. Qualitative Results (Presence or Absence)
Limit of detection
Selectivity determined to ensure interference not occurring-use a range of micro-organisms

34. Quantitative methods Sensitivity Selectivity Precision
Expression of uncertainty

35. Environment Restricted access Lab coats
‘Potential for contamination minimised’ ie non-absorbent surfaces,easy clean, blinds,no plants,no personal possessions,minimal paperwork
Documented cleaning programme

36. Cont Microbiological monitoring Records kept Action and alert limits
Documented procedures

37. Sample/Deposit Handling and Identification
System can be simple as notebook or complex LIMS system
Adequate facilities for storage and segregation essential (fridges,freezers)
Samples must be stored and examined under conditions which minimise changes in the numbers or viability of micro-organisms

38. Disposal of Contaminated Waste
All waste should be autoclaved before leaving the laboratory
If disposal of waste is subcontracted control measures must be in place
However if pathogens involved also follow ACDP guidelines and WHO Manuals

39. Organisms in Quality Systems
Use, preservation and traceability

40. Organisms in Quality Systems
Challenge testing
Method validation
Media QC
Analytical Quality Control (AQC)
Proficiency Testing

41. Guidance for ISO 17025 States two fundamental requirements:-
‘…shall maintain records of checks on batch to batch variation of media and other reagents by using working standards of known micro-organisms’
‘...shall hold reference cultures obtained from a recognised national collection in order to demonstrate traceability’

42. Common Confusions Definitions of reference/stock/working cultures
Can you subculture and if so how many times
Are the newer delivery systems acceptable

43. Reference Organisms-1
Reference cultures (i.e. from a collection) may be subcultured once to provide reference stocks
Reference stocks shall be maintained ‘...by techniques which maintain the desired characteristics of the strains’
Examples-freeze drying, LN,deep freeze methods

44. Reference Organisms-2
Reference stocks are used to prepare working stocks for routine use
Working stocks can be subcultured if:-
required by a standard method
documentary evidence exists to show changes do not occur over a defined number of subcultures, and this has been validated

45. Reference Organisms-3
Once thawed or otherwise reconstituted reference stocks must not be refrozen or re-used
Working stocks must not be sub-cultured to replace reference stocks

46. Things often forgotten
Document traceability of strains including date purchased,dates sub-cultured dates beads removed etc..
Continuous monitoring of freezer temp. or at least max/min.
Place shelf life on all stocks, reference and working

47. ‘New’ Delivery Systems
E.g Culti loops, Quanticult etc
All previous requirements still apply
Proof of quality, traceability etc falls to the laboratory
Treat like any other goods and service supply but with extra ‘calibration’ requirements

48. Questions to Ask ISO 9000.. Company?
But what for -manufacture,supply? Who actually makes the products?
Traceability to national collection?
How many passages?
What preservation techniques are used?

49. Documents Required Validated methods and QC data
Certificate of traceability to National Collection number
Statement of uncertainty i.e. if quantitative, number of cfu +/- but also details of how this is calculated, cumulative errors etc.

50. Finally
Whatever route chosen, the onus falls on the laboratory to demonstrate quality is being maintained.

51. Media Quality Control
Why and How

52. Media QC
Checked to ensure support growth of appropriate organisms (this includes negative control)
Must be quantitative if counts involved
Sterility must be confirmed
pH,and other parameters set by manufacturer must be checked post sterilisation

53. Diluent QC As media where appropriate
Volume of batches dispensed prior to autoclave must be established post sterilisation and monitored at intervals if stored
Select percentage, assess gravimetrically

54. Media QC Records- Type Batch number Container pH
Date (of set up and read)
Sterility
Volume
Test strains (+ve,-ve)
Test conditions (temp,atmosphere)
Results (expected and actual)
Signature of staff
Cross ref to autoclave batch and strain records

55. QC Methods Simple streak/assess growth and morphology
Quantitative/known count
Spiked simulated samples-available for purchase or prepare in-house

56. References
Brown,MRW;Gilbert P. Microbiological Quality Assurance: a guide towards relevance and reproducibility. Boca Raton,FL:CRC Press, 1994
National Committee for Clinical Laboratory Standard M22-A2. Quality Assurance for Commercially prepared microbiological culture media. Wayne PA NCCLS, 1996