Proficiency testing (PT) scheme for Shellfish. ISO what does compliance as a PT provider mean?

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1 Proficiency testing (PT) scheme for Shellfish ISO what does compliance as a PT provider mean? Nita Patel Food and Environmental Proficiency Testing Unit 10 November 2016

2 Contents Public Health England s mission for National Infection Service About Shellfish testing About proficiency testing Why laboratories do PT ISO and compliance Summary of the benefits

3 Mission National Infection Service (NIS) The mission and core purpose of PHE for NIS is to protect the population of England from infectious disease. It does this by: a) Detecting outbreaks and monitoring communicable disease; b) Analysing data collected on communicable diseases and their causes, providing outbreak detection tracking and national surveillance; c) Researching and effecting interventions, both immediately (emergency response to outbreaks and incidents as a Category 1 responder, defined under the 2004 Civil Contingencies Act) and through longer-term prevention; d) Providing expert scientific advice and guidance to the public, government and international agencies

4 Introduction The consumption of sewage polluted bivalve molluscan shellfish (BMS) may cause illness in the consumer through ingestion of faecally borne pathogens (such as Salmonella, norovirus and hepatitis A virus) The risks of exposure to these infectious agents are compounded by the traditional consumption of raw, or only lightly cooked BMS. Enteric bacteria, such as Escherichia coli are used as indicator organisms to assess the sanitary quality of shellfish and their growing waters and to predict the risk of exposure to enteric pathogenic microorganisms

5 Public Health concerns Shellfish areas often impacted by sewage Bivalves can filter litres per hour Pathogens from sewage are concentrated (~100x) Shellfish are eaten raw or lightly cooked Outbreaks of disease can occur on large scale

6 European Union Regulation 854/2004 In the European Union (EU), the criteria for laying down the microbiological standards for BMS are set out in Regulation (EC) 854/2004 and Regulation (EC) 2073/2005. This stipulates the conditions for the production and placing on the market of live BMS Across the EU E. coli in shellfish flesh is used as an indicator of faecal contamination of BMS The EU reference method for the determination of E. coli in BMS is ISO * *ISO Microbiology of food and animal feeding stuffs Horizontal method for the enumeration of β glucuronidase positive Escherichia coli Part 3: Most probable number technique using 5 bromo 4 chloro 3 indolyl β D glucuronide

7 Roles and Responsibilities England and Wales Food Standards Agency Competent Authority Makes final classification decisions Issues classification list Determines policy

8 Facts and Figures - UK Scale of programme: 75 production areas 375 beds classified: oysters, mussels, cockles and various clams Approximately 90,000 results

9 Facts and Figures - UK Scale of programme: 75 production areas 375 beds classified: oysters, mussels, cockles and various clams Approximately 90,000 results There is a need for examination of samples using approved method/s and accurate reporting of results to ensure that public health occurrences can be effectively managed

10 Food and Environmental Proficiency Testing Unit (FEPTU)

11 External quality assessment (EQA) / Proficiency Testing (PT) explained Samples of known but undisclosed content are introduced into a laboratory s routine testing procedure Freeze-dried LENTICULE discs

12 Proficiency Testing ORGANISING LABORATORY PARTICIPANTS Prepare samples Examine samples Analyse results Report results Prepare report Evaluate

13 Overview - Why laboratories do PT To demonstrate competence as part of your accreditation requirement ISO/IEC 17025: General requirements for the competence of testing and calibration laboratories Helps to provide assurance of the results obtained provided they are treated and processed the same as other samples Helps improve laboratory processes and understanding of regulation/legislation To remain up to date with new and emerging organisms - educational To challenge processes/media/training with difficult or atypical organisms Inter-laboratory comparison of performance To support work tendered for as an accredited laboratory Because you enjoy the challenge that participating in PT brings!

14 Impact of incorrect results food microbiology E.coli levels too low or false negative Salmonella result: Unsafe product released for sale People become ill/die Negative publicity Loss of reputation Financial cost E. coli levels too high or false positive Salmonella result: Incorrect classification Unnecessary disruption Restriction or ban on the export of the product Loss of confidence in harvesting area Financial cost 14

15 Laboratory mistakes Incorrect results may arise from: Sample handling errors (mixed up, incorrectly labelled etc.) Calculation errors and misreading the most probable numbers (MPN) tables Cross-contamination Inadequate staff training Equipment and culture media failures Reporting errors

16 Shellfish Scheme This scheme provides external quality assessment samples for laboratories that examine raw bivalve molluscs from harvesting sites in accordance with Regulation (EC) No. 854/2004 and from the production chain between harvest and consumption, in accordance with Regulation (EC) 2073/2005 as amended. The Scheme is organised in collaboration with the Centre for Environment, Fisheries and Aquaculture Science (Cefas), Weymouth, United Kingdom (UK). The scheme focuses on raising awareness: of interpreting tube combination results and the associated MPN value per 100g for Escherichia coli for classification of harvest beds of problems with Salmonella detection (included in the scheme for end-product testing, not for classification purposes) of updates in the ISO method

17 Shellfish Scheme Three distributions a year with two samples in each (2 labs in Peru) A total of 63 participants in 30 different countries Mandatory participation for the National Reference Laboratories (NRL) in each member state reporting to the European Union Reference Laboratory (EURL - Cefas) Permission from the NRLs is obtained to release performance date to the EURL

18 Most Probable Numbers (MPN) The method used to determine the MPN has greater inherent variability than colony count methods However, it is sometimes required for the detection and estimation of levels when low levels of micro-organisms are expected, especially when the microorganisms may be stressed (e.g. as a result of processing, freezing etc.) The MPN method is stipulated in European Union criteria for the microbiological examination of certain products such as dairy products and live bivalve molluscs and other shellfish Most results are reported as the number of positive tubes out of 5 at 3/4 dilutions and the corresponding most probable number (MPN) (electronic calculator)

19 How do you measure performance?

20 Statistics used Participating in PT is the only way a laboratory can obtain an ongoing independent assessment of its performance To get the maximum benefit from participating in PT it is helpful to know how the statistics that are used are applied FEPTU uses statistics to determine whether a participants results are within an expected range

21 Statistics used The expected range comes from the distribution (average and variability) of all the participants results the median value ISO 13528: Statistical methods for use in proficiency testing by interlaboratory comparisons is followed Certain criteria must be followed when analysing participants results

22 Setting the criteria The statistical design for PT schemes for microbiology are influenced by: Homogeneity level in a sample (distribution of organisms) Whether the test is for the detection or the enumeration of a target organism Number of participants in a scheme Target level of organism Method of analysis For example, results for an examination using colony count methods will be analysed in a different way to those where determination of the most probable number (MPN) is required

23 Scheme Reports Summary of all participants results Assessment of every result using a scoring system Assessment of each participant s performance over time Scatter graphs to help participants compare performance with other laboratories Advice and guidance

24 Summary of all participants results

25 Scheme Reports Summary of all participants results Assessment of every result using a scoring system Assessment of each participant s performance over time Scatter graphs to help participants compare performance with other laboratories Advice and guidance

26 PHE scores applied E. coli MPN result is determined from the median result reported by participants ± 2.68 standard deviations (SD). The SD values used to determine the expected range take into account the expected inherent variability of the three by five tube MPN method, which on a log 10 scale has a value of 0.26 Results are defined as follows: A) Expected range = median ± 2.68 SD B) Outlying result 1 = median ± >2.68 ± 4 SD C) Outlying result 2 = median ± >4 SD Marks are deducted if tube combination is inconsistent with MPN reported

27 Scheme Reports Summary of all participants results Assessment of every result using a scoring system Scatter graphs to help participants compare performance with other laboratories Assessment of each participant s performance over time Advice and guidance

28 Scatter graph of results

29 Scatter graph of results

30 Scheme Reports Summary of all participants results Assessment of every result using a scoring system Scatter graphs to help participants compare performance with other laboratories Assessment of each participant s performance over time Advice and guidance

31 On going performance over time Is designed to alert participants with on-going problems Is only useful if a laboratory takes more than one distribution a year in a scheme Allows laboratories to identify errors in the system Is an effective tool for on-going monitoring of quality control processes Outlying results are allocated a low score On single occasion reporting an outlying result participants must not be alarmed but they should access the reasons why Laboratories that achieve less than 70% of the maximum possible score are likely to be experiencing significant problems

32 On-going performance data - table

33 On-going performance data - graph

34 Trend analysis Trend analysis tools are available on our web-site, these help participants to plot their own results to identify performance trends against the participants median. See example below: Trend analysis tools are available under each scheme heading:

35 ISO compliance

36 ISO versus ISO 17025

37 ISO versus ISO 17025

38 ISO versus ISO 17025

39 ISO introduction

40 ISO some issues It is a very generic document written mainly by chemists The language used does not always make sense for microbiological criteria For microbiology we need to take into account stability, variability and homogeneity, behaviour of the organisms etc. To address specific requirements for microbiology ISO/TS 22117:2010 was published (Microbiology of food and animal feeding stuffs - Specific requirements and guidance for proficiency testing by interlaboratory comparison)

41 ISO critical aspects

42 ISO Planning (4.4.1) The proficiency testing provider shall not subcontract the planning of the proficiency testing scheme The proficiency testing provider shall document a plan before commencement of the proficiency testing scheme

43 ISO Homogeneity and stability (4.4.3) Criteria for suitable homogeneity and stability shall be established and shall be based on the effect that inhomogeneity and instability will have on the evaluation of the participants' performance. Homogeneity organisms are distributed in a sample Stability the levels of the organisms are similar over a defined period of time For the Shellfish Scheme difficult to prove homogeneity due the inherent variability of the MPN method. Method for assessing participants results assumes that the sample is well-mixed prior to testing and therefore allows for this variability (wider standard deviation used)

44 ISO Homogeneity and stability (4.4.3) FEPTU Quality Control: To demonstrate stability of the sample, a minimum of ten LENTICULE discs, selected randomly from a batch, are examined throughout the distribution period for E. coli enumeration and for Salmonella detection

45 ISO Statistical design (4.4.4) Statistical designs shall be developed to meet the objectives of the scheme, based on the nature of the data (quantitative or qualitative, including ordinal and categorical), statistical assumptions, the nature of errors, and the expected number of results The proficiency testing provider shall document the statistical design and data analysis methods to be used to identify the assigned value and evaluate participant results, and shall provide a description of the reasons for their selection and assumptions upon which they are based In designing a statistical analysis, the proficiency testing provider shall give careful consideration to the following: accuracy, number of participants, identify and how to handle outliers, scoring criteria etc.

46 ISO Statistical design (4.4.4) PHE statistician is consulted on all unusual results reported by participants

47 ISO Packaging, labelling and distribution of PT items (4.6.3) The proficiency testing provider shall control packaging and labelling processes to the extent necessary to ensure conformity with relevant national, regional, or international safety and transport requirements The proficiency testing provider shall specify relevant environmental conditions for the transport of proficiency test items. Where relevant, the proficiency testing provider shall monitor the pertinent environmental conditions of the proficiency test item during transport and assess the impact of environmental influences on the proficiency test item

48 ISO Packaging, labelling and distribution of PT items (4.6.3) This ensures that everybody receives samples that are the same across the world We therefore examine samples as part of our QC process that have gone through the shipment process to identify impact on the sample contents and levels

49 ISO Evaluation of performance (4.7.2) The proficiency testing provider shall use valid methods of evaluation which meet the purpose of the proficiency testing scheme. The methods shall be documented and include a description of the basis for the evaluation. The evaluation of performance shall not be subcontracted Where appropriate for the purpose of the proficiency testing scheme, the proficiency testing provider shall provide expert commentary on the performance of participants

50 ISO Evaluation of performance (4.7.2)

51 ISO Reports (4.8.1) Proficiency test reports shall be clear and comprehensive and include data covering the results of all participants, together with an indication of the performance of individual participants. The authorization of the final report shall not be subcontracted Reports shall be made available to participants within planned timescales - this allows for early investigation of possible error

52 ISO Reports (4.8.1)

53 ISO Reports (4.8.1)

54 ISO Reports (4.8.1)

55 ISO Reports (4.8.1)

56 ISO Reports (4.8.1)

57 ISO Reports (4.8.1)

58 ISO Confidentiality (4.10) The identity of participants in a proficiency testing scheme shall be confidential and known only to persons involved in the operation of the proficiency testing scheme, unless the participant waives confidentiality

59 Why participate? PHE PT samples are designed to challenge your testing procedures therefore will include challenging organisms so beware We extensively test the samples using ISO methods so your results should align with our results including confirmatory test results Process PT samples the same as other routine samples. Otherwise nothing will be learnt about your quality system We are not here to trick you but to: raise awareness of the limitation/s of your procedure or method provide an insight into staffs knowledge and experience endorse the requirement to carry out confirmatory tests encourage the use of approved methods give you an opportunity to examine samples containing organisms less frequently encountered that are of public health concern

60 Summary of the benefits to you Raises awareness of new and emerging organisms of public health concern Provides an opportunity to improve staffs knowledge and experience with organisms not frequently encountered Better understanding of your performance in relation to the method/s you use Greater understanding of the limitation/s of your method especially with atypical/emerging organisms Highlights impact on results when approved methods are not followed Highlights impact on results with unusual, atypical strains or less commonly encountered organisms Gives you an independent assessment of your laboratory s overall performance Allows you to identify gaps in your process where quality improvements are required

61 Other schemes available are Water PT Legionella Isolation Scheme Including one specific for molecular Drinking Water Scheme Recreational and Surface Water Scheme Bathing beach (marine) Swimming pool River, lakes and streams Bottle and Mineral Water Scheme Endoscope Rinse Water Scheme Dialysis Water Scheme Hospital Tap Water Scheme Food PT Standard Scheme European Food Microbiology Legislation Scheme Non-Pathogen Scheme Shellfish Scheme Pathogenic Vibrio Scheme Staphylococcus aureus Enterotoxin Scheme Environmental Swab Scheme Hygiene indicator Pathogen detection Shiga toxin E. coli molecular

62 Acknowledgements Angela Appea Richard Borrill Kermin Daruwalla Joanna Donn Carmen Gomes Thomas Harper Salah Himedan Zak Prior Manchari Rajkumar Aneta Stranc Lili Tsegaye Vanessa Waite PHE Steering Group members eqad clinical team Andy Younger (Cefas) Louise Stockley and Cefas team

63 Thank you for listening

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