Quality Management -Standards Joan Kelley 28th Nov 2002

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

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

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

Context Regulations = EN45001, European Standard (others GLP, ISO 9000 etc) But since early 2000…………. ISO/IEC 17025 came into force General Requirements for the Competence of Testing and Calibration Laboratories

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

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

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

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

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

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

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

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

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

Application of Standards to Microbiology labs Some Guidelines

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)

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

Quantitative methods Sensitivity Selectivity Precision Expression of uncertainty

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

Cont Microbiological monitoring Records kept Action and alert limits Documented procedures

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

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

Organisms in Quality Systems Use, preservation and traceability

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

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’

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

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

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

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

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

‘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

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?

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.

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

Media Quality Control Why and How

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

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

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

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

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