HISTOCOMPATIBILITY. and IMMUNOGENETICS. Prospectus

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1 HISTOCOMPATIBILITY and IMMUNOGENETICS Prospectus 2014

2 CONTENTS Page 1. Distribution Timetable 2 2. Confidentiality 2 3. Participation Registration Service s Expectations Guidance on Participation Bespoke Schemes Laboratory Performance Reports and Tables Charges for Participation 5 4. UK NEQAS for H&I Assessment and Performance Principles 5 5. Nomenclature 6 6. Scheme 1A - HLA Phenotyping 7 7. Scheme 1B - HLA-B27 Testing 9 8. Scheme 2A - Cytotoxic Crossmatching Scheme 2B - Crossmatching by Flow Cytometry Scheme 3 - HLA Antibody Specificity Analysis Scheme 4A1 DNA HLA Typing at 1 st Field Resolution Scheme 4A2 DNA HLA Typing to 2 nd Field Resolution Scheme 4B - ABO Grouping by DNA-based Methods Scheme 5A - HFE Typing Scheme 5B Interpretative: HFE Genotype and Hereditary Haemochromatosis Scheme 6 HLA Antibody Detection Scheme 7 HLA-B*57:01 Typing for Drug Hypersensitivity Scheme 8 HLA and Disease Typing for HLA-DR/DQ/DP only Educational Scheme Essential Scheme Information UK NEQAS for H&I Steering Group Prospectus HLA Nomenclature Test Material UK National Quality Assurance Advisory Panel (UK NQAAP) Annual Participants Meeting Endorsement by UK NEQAS for H&I Scheme Review and Pilot Schemes Procedure for the Handling of Complaints from Participants Appeals Against an Assessment Non-Analytical Errors Correspondence with Participants Policy on Testing Returned Scheme s Material Acceptance of Participants Results Contact with UK NEQAS for H&I List of Contact Names and Addresses Joint Working Group for Quality Assurance Guidelines for Participants CPA Accreditation and EQA Performance UK NEQAS for H&I and European Federation for Immunogenetics (EFI) Accreditation UK NEQAS for H&I Dates 36 1

3 PROSPECTUS DISTRIBUTION TIMETABLE Scheme 1A Scheme 1B Scheme 2B Scheme 3 Scheme 4A1/4B Scheme 4A2 Scheme 8 Scheme 2A Scheme 6 Scheme 5A Scheme 7 Educational Scheme Scheme 5B 21 January 7 January 14 January 28 January 11 February 18 February 11 February 25 March 4 March 11 March 13 May 20 May 3 June 24 June 17 June 10 June 17 June 15 July 8 July 1 July 16 September 9 September 23 September Scheme 1A - HLA Phenotyping Scheme 1B - HLA-B27 Testing Scheme 2A - Cytotoxic Crossmatching Scheme 2B - Crossmatching by Flow Cytometry Scheme 3 - HLA Antibody Specificity Analysis Scheme 4A1 DNA HLA Typing at 1 st Field Resolution Scheme 4A2 DNA HLA Typing to 2 nd Field Resolution Scheme 4B - ABO Grouping by DNA-based methods Scheme 5A - HFE Typing Scheme 5B Interpretative: HFE Genotype and Hereditary Haemochromatosis Scheme 6 HLA Antibody Detection Scheme 7 HLA-B*57:01 Typing for Drug Hypersensitivity Scheme 8 HLA and Disease Typing for HLA-DR/DQ/DP only Educational Scheme samples and Interpretive Scenarios 2. CONFIDENTIALITY Laboratory code information is known only to the Schemes Director, Manager and UK NEQAS for H&I staff. Laboratory identifiers and performance information are confidential and will not be released to a third party without the written permission of the Head of the participating laboratory. However, for UK laboratories, unsatisfactory performance will be notified to the Chairman and Members of the UK National Quality Assurance Advisory Panel (UK NQAAP) for Immunology and laboratories may be identified to the Chairman of the Joint Working Group for Quality Assurance (JWG) (see Section 23). UK laboratories identified to UK NQAAP for Immunology regarding Scheme 5A will also be identified to the UK NQAAP for Clinical Cytogenetics and Molecular Genetics. 3. PARTICIPATION 3.1 REGISTRATION Registration forms are provided before the commencement of the UK NEQAS for H&I year. By signing these registration forms you agree to abide by the JWG Conditions of EQA Scheme Participation (Section 23) Guidelines (UK laboratories only) and the list of Service Expectations below. You also agree to pay the fees and any agreed courier charges in a timely manner. 3.2 SERVICE S EXPECTATIONS Our commitment to UK NEQAS for H&I Participants we will: Respect your confidentiality Despatch samples according to the published timetable Accurately maintain our contact database Not give participant information to anyone (except as detailed in the Prospectus) Provide you with all the information for you to fully participate in our schemes Provide schemes at cost and will not make a profit Rectify assessment errors in a timely fashion Resolve disputes in an impartial and professional manner Willingly provide advice on all scheme issues Endeavour to comply with EFI EPT Standards for Providers 2

4 Your commitment to UK NEQAS for H&I you will: Notify us if you do not receive the expected samples Test and interpret EQA samples as clinical specimens Always give the reason(s) for not testing a NEQAS sample Provide up-to-date contact information Not share scheme findings with other laboratories until after our report has been issued Abide by the JWG Conditions of EQA Scheme Participation (UK laboratories only) 3.3 GUIDE TO LABORATORY TESTING/CLINICAL SERVICES AND SCHEME PARTICIPATION Participation in a particular UK NEQAS for H & I scheme is at the discretion of individual laboratories. As a rule, laboratories should take part in external quality assessment (EQA) schemes that reflect, as far as possible, their clinical testing practices. The simplest example is the operation of a clinical service for HLA-B27 typing and participation in Scheme 1B (HLA-B27 Testing). UK NEQAS schemes are generally analyte, rather than technique, driven. Thus, again in the simplest situation, if clinical HLA-B27 testing is offered as a clinical service, laboratory participation would be in Scheme 1B whether the method used was cell/antibody or DNA-based. However, UK NEQAS for H & I schemes have evolved by attempting to take into account changing technologies and clinical practices. The obvious example of this is Scheme 2 where 2A assesses the outcome of cell/serum cross-matching by lymphocytotoxicity-based techniques and 2B by flow cytometry. The following Table provides a guide to Scheme participation: Examples of Laboratory Testing/Clinical Services: Full HLA typing in: Solid organ transplantation Haematopoietic stem cell transplantation HLA and disease investigations Unrelated haematopoietic stem cell donor registries Reference cell panel typing for clinical HLA antibody services HLA-B27 testing in: Aid to diagnosis in B27 associated diseases ABO blood grouping (using DNA) in: Solid organ transplantation Typing for single specific HLA specificity/allele or locus, other than HLA-B27 in: Disease susceptibility/aid to diagnosis Immunotherapy HLA antibody detection and specification in: Transplantation Platelet immunology Blood products support Crossmatching by lymphocytotoxicity in: Transplantation Blood products support Crossmatching by flow cytometry in: Transplantation HFE testing in: Patient investigations Haemochromatosis family studies Drug hypersensitivity testing in: Patient evaluation Participation in Scheme: 1A and/or 4A1 and/or 4A2 1B 4B 1A and/or 4A1 and/or 4A2 and/or 8 3 or 3 and 6 2A 2B 5A and /or 5B 7 PARTICIPATION IN SCHEME 1A (HLA PHENOTYPING) AND SCHEMES 4A1/4A2 (DNA HLA TYPING) The conduct of Schemes 1A and 4A1/4A2 has attempted to keep reasonable pace with changing HLA testing technology and typing strategies. Modifications to the participation levels of these Schemes have tried to encompass these changes. 3

5 In view of the increasing complexities of HLA typing methods, strategies and services the following guidelines are intended to help laboratories in their decision to participate in Scheme 1A (HLA Phenotyping) and/or Schemes 4A1/4A2 (DNA HLA Typing). Scheme 1A is aimed at laboratories undertaking HLA-A, B, C, DR, DQ typing or any combination of these, by serology. Participants must only register to be assessed on those loci tested using serological methods. The Steering Committee acknowledges that many laboratories use supplementary techniques to aid in the definition and refinement of HLA specificities detected by serology. Reporting and assessment must use HLA specificity nomenclature. Based on previous years practice there will ordinarily be an expectation that the broad HLA specificities, e.g., B15, B40, DR3 and DQ3 will be subdivided into their component split specificities. Schemes 4A1/4A2 (DNA HLA typing) will be performed by laboratories that undertake DNA based typing and, typically, might routinely perform a combination of 1 st field typing (formerly known as low resolution or 2-digit typing) and 1 st and 2 nd field typing (formerly known as high resolution or 4-digit typing). For example, 1 st field HLA-A, and B typing and 1 st and 2 nd field DRB1 typing. For this reason it is possible to register for any combination of HLA-A, B, C, DRB1, DRB3, DRB4, DRB5, DQA1 and DQB1 for 1 st field typing or 1 st and 2 nd field typing and DPB1 for 2 nd field typing and provide data for DPA1 for comparison purposes. Registrants for 1 st field typing can also sign up for reporting merely the presence of DRB3/4/5. The European Federation for Immunogenetics (EFI) STANDARDS FOR HISTOCOMPATIBILITY & IMMUNOGENETICS TESTING version 6.1, D - HLA ALLELES AND ANTIGENS, Standard D1.4 states: High resolution typing is defined as the identification of HLA alleles that encode the same protein sequence within the antigen binding site. HLA alleles must be identified at the level of resolution which defines the first and second fields according to WHO nomenclature by at least resolving all ambiguities: resulting from polymorphisms located within exons 2 and 3 for HLA class I loci, and exon 2 for HLA class II loci that encompass a null allele, wherever the polymorphism is located, unless it can be demonstrated that an expressed antigen is present on the cells. It is expected that laboratories that register for HLA typing to the 2 nd field will have developed a strategy to comply with this Standard with regard to the recognition of null alleles. The following Table provides a guide to HLA typing practices and the most applicable HLA typing schemes. HLA typing practice Scheme(s) Serology 1A Serology with 1 st field DNA typing 1A or 1A and 4A1 1 st field DNA typing 4A1 Combination of 1 st field and 2 nd field DNA typing 4A1 and 4A2 2 nd field DNA typing 4A2 PARTICIPATION IN SCHEME 3 (HLA ANTIBODY SPECIFICITY ANALYSIS) AND SCHEME 6 ( HLA ANTIBODY DETECTION) Most H & I laboratories provide a service for testing of patients sera for the presence of antibodies directed towards HLA-class I (A, B, C) or class I and class II (DR, DQ, DP) specificities. This may be within the context of solid organ or haematopoietic stem cell transplantation, provision of blood products or the investigation of a clinical condition, e.g. thrombocytopaenia. Many laboratories undertake this service using a two-tier system. The first stage antibody detection determines which sera should be selected for the comprehensive second stage antibody specificity assignment. Both stages are equally important since failure to detect the presence of antibodies during screening will obviously deny that serum the benefit of the often more comprehensive testing associated with antibody specification. However, where numerous patients sera are tested an effective two-stage system can significantly reduce laboratory workload thus allowing more resources to be applied to the important antibody specificity assignment stage. The test sera supplied in Scheme 6 (HLA Antibody Detection) may or may not contain HLA antibodies directed towards HLA-class I and/or class II specificities. The purpose of this Scheme is to assess the laboratory s ability to undertake the initial screening process which places individual patients sera into the no HLA antibodies detected category or HLA antibodies probably present requires further analysis category. The method(s) used should be those routinely employed by the laboratory for antibody testing. 4

6 For Scheme 3 (HLA Specificity Analysis) sera are provided that are known to contain antibodies directed towards HLA-class I and/or class II specificities. The purpose of this Scheme is to assess the laboratory s ability to determine the component HLA specificity or specificities in the antisera using all methods the laboratory employs for clinical samples. PARTICIPATION IN SCHEME 7 HLA-B*57:01 TYPING FOR DRUG HYPERSENSITIVITY Pharmacogenetics, generally accepted as the study, or clinical testing, of genetic variation that gives rise to differing responses to drugs, is increasingly being applied to the prospective testing of patients for HLA-B*57:01 who are to be treated with the antiretroviral drug abacavir. Thus, a strong association exists between HLA-B*57:01 and the development of drug hypersensitivity reactions to abacavir. This scheme tests participants ability to define HLA-B*57:01 using the method(s) they employ for routine clinical service HLA-B*57:01 testing. PARTICIPATION IN SCHEME 8 HLA AND DISEASE TYPING FOR HLA-DR/DQ/DP ONLY This scheme, which uses selected DNA extracts, is aimed at laboratories that provide a service as an aid to the diagnosis of certain HLA Class II-associated diseases, e.g. actinic prurigo, coeliac disease and narcolepsy. 3.4 BESPOKE SCHEMES UK NEQAS for H&I is able to provide specific EQA schemes for certain HLA and HFE alleles Bespoke schemes will use selected stored DNA from samples previously tested in UK NEQAS for H&I s Schemes. Consequently, these samples are considered as well-documented reference material. Provision of a bespoke scheme also applies to HLA antibody detection and/or specification. A further use of bespoke schemes is fulfilling the requirements of EFI Standard C6.4 If a laboratory's performance in EPT programme(s) is unsatisfactory in any category for which EFI accreditation is sought, the laboratory must: participate in an additional EPT programme in that category and document the Director s review and any corrective action taken. Due to the nature of Scheme 2A - Cytotoxic Crossmatching and Scheme 2B - Crossmatching by Flow Cytometry, UK NEQAS for H&I are normally unable to supply additional sera/blood samples for these Schemes. Requests for a bespoke scheme, giving full details of requirements, should be made to: Deborah Singleton (Schemes Manager, UK NEQAS for H&I, Welsh Blood Service, Ely Valley Road, Talbot Green, Pontyclun CF72 9WB. Tel: ; Fax: ; LABORATORY PERFORMANCE REPORTS AND TABLES Individual laboratory performance reports and performance tables will be posted to participants. Results summaries may be downloaded from the website: Excel spreadsheets of Scheme 3 results are available on request from: 3.6 CHARGES FOR PARTICIPATION These are shown on the insert sheet. For further information please contact: Deborah Singleton, Schemes Manager, UK NEQAS for H&I, Welsh Blood Service, Ely Valley Road, Talbot Green, Pontyclun CF72 9WB. Tel: ; Fax: ; 4. UK NEQAS FOR H&I ASSESSMENT AND PERFORMANCE PRINCIPLES This information should be read in conjunction with each Scheme s ASSESSMENT and SATISFACTORY PERFORMANCE sections. Scheme assessment and satisfactory performance criteria are reviewed by the Steering Committee each year and take into account current EFI EPT Standards for Providers established by the EFI EPT Committee. Proposed changes in Schemes assessment and/or the determination of satisfactory performance are brought to the UK NQAAP for Immunology for ratification. For all Schemes, except Scheme 5B - Interpretative: HFE Genotype and Hereditary Haemochromatosis, performance is based on establishing consensus assignments, for HLA/ABO/HFE specificities/alleles, antibody presence/absence, antibody specificities, cell/serum crossmatch test positive/negative, for all test samples. Scheme 5B uses a penalty points system. 5

7 The general consensus level for assignments is 75%, thus assignments reaching a 75% or greater agreement between participants will be assessed while those of <75% agreement are not assessed. Scheme 3 - HLA Antibody Specificity Analysis - also uses a 95% consensus level for the absence of an HLA specificity. The actual assessment procedure is detailed for each scheme under Assessment Procedure. In general reporting a consensus assignment is considered as Acceptable Performance while reporting a non-consensus assignment is considered Unacceptable Performance. Satisfactory performance is generally based on achieving a specified number of sample (patient) reports in agreement with consensus assignments in a calendar year. Satisfactory performance levels are set according to those established by EFI (minimum performance standards) or greater. For Schemes involving the assignment of a single HLA specificity/allele, an ABO group or an HFE type the Satisfactory Performance level is set at achieving all the correct consensus assignments in all samples supplied during one calendar year. For other schemes, some leniency regarding full compliance with consensus assignments is allowed (detailed for each scheme under Satisfactory Performance ). Laboratories not achieving Satisfactory Performance will receive written notification of their Unsatisfactory Performance status, as soon as it occurs, and will be expected to detail their reasons for their performance together with any corrective actions within 20 working days of the date of the Unsatisfactory Performance notification. Note: this does not apply to Interpretative Schemes, e.g. Scheme 5B. All UK laboratories receiving Unsatisfactory Performance notification are reported to UK NQAAP for Immunology. Failure to reply within 20 working days of the date of an Unsatisfactory Performance notification or replies deemed unsatisfactory will be specifically reported to UK NQAAP for Immunology who may take further action. 5. NOMENCLATURE Participants are requested to report their findings using the correct nomenclature relevant to the methodology used. The most recently published FULL WHO Nomenclature Report will be taken as the reference baseline nomenclature; i.e. participants will be expected to report their findings in accord with this report as a minimum. HLA Nomenclature Reports and HLA Dictionary are available from: In addition, the Steering Committee has considered the reporting of HLA alleles and has formulated the following convention for reporting groups of alleles: Please use the published up-to-date HLA nomenclature, see above. Please report alleles fully, e.g. DRB4*01:02-01:03 and not as DRB4*01:02-03 which may be interpreted as DRB4*01:02-01:03 or as DRB4*01:02-03:01. Groups of alleles should be reported as allele x / allele y, where / means or, e.g. DRB1*01:01/01:02/01:04 means DRB1*01:01 or DRB1*01:02 or DRB1*01:04. Groups of alleles that include sequential allele numbers may be reported as allele x - allele y, where - means to, e.g. DRB1*15:01-15:04 means that the allele in question could be any of the alleles between DRB1*15:01 to DRB1*15:04 inclusive, i.e. DRB1*15:01 or DRB1*15:02 or DRB1*15:03 or DRB1*15:04. Please report DRB5*, DRB3* and DRB4* and not their serological equivalents, i.e. DR51, DR52 and DR53. 6

8 SCHEME 1A 6. SCHEME 1A HLA PHENOTYPING PURPOSE To assess participants ability to use serological and supplementary methods to correctly identify HLA specificities. 6.1 SAMPLES A total of ten blood samples will be sent each year as five distributions of two blood samples. 6.2 REPORTING Depending on their typing strategies participants may register for HLA Class I typing only or for HLA Class I and II typing Participants must only register to be assessed on those loci tested using serological methods Participants must make a report for each HLA locus for which they have registered It is acknowledged that many laboratories use supplementary techniques to confirm serological HLA specificity assignment The report should detail the HLA phenotype using official WHO HLA specificity nomenclature Reports should be made at the split specificity level, where appropriate, using the results of the serological typing and any typing performed using supplementary techniques Participants must only use the reporting forms provided Participants are required to make their report within 10 days. 6.3 ASSESSMENT Participants will be assessed on the HLA loci contained in the phenotype, for which they are registered The consensus complete HLA phenotype for assessment is determined by at least 75% of laboratories agreeing each specificity Specificities failing to reach the 75% consensus level will not be assessed A "blank" forms part of the assessment if at least 75% of laboratories report a single specificity at a locus Assessment Procedure Each complete HLA phenotype in agreement with the consensus phenotype Each complete HLA phenotype not in agreement with the consensus phenotype Acceptable Unacceptable Each sample report of a not tested result see SATISFACTORY PERFORMANCE Satisfactory performance is obtaining nine or more complete HLA phenotypes in agreement with the consensus phenotypes in a calendar year Laboratories with unsatisfactory performance will receive written notification of their status and will be expected to reply to UK NEQAS for H&I detailing their corrective actions For UK laboratories, unsatisfactory performance will be reported to UK NQAAP for Immunology For UK laboratories, failure to reply or replies deemed unsatisfactory are likely to be further actioned by UK NQAAP for Immunology. 7

9 SCHEME 1A 6.5 INFORMATION/ANALYSIS PROVIDED FOR PARTICIPANTS Summary sheets of all reported specificities and supplementary information Information on methodology Performance information on the current and previous year s samples for all laboratories. 8

10 SCHEME 1B 7. SCHEME 1B - HLA-B27 TESTING PURPOSE To assess participants ability to correctly determine HLA-B27/B2708/B*27 status. 7.1 SAMPLES A total of ten blood samples will be sent each year as five distributions of two blood samples. 7.2 REPORTING Participants are required to report on HLA-B27/B2708/B*27 positivity or negativity Participants must only use the reporting forms provided Participants are required to make their report within 21 days. 7.3 ASSESSMENT The HLA-B27 status of each sample is determined by at least 75% of laboratories agreeing on the presence or absence of HLA-B Samples failing to reach the 75% consensus level will not be assessed Assessment Procedure Each sample report in agreement with the consensus HLA-B27 status Each sample report not in agreement with the consensus HLA-B27 status Acceptable Unacceptable Each sample not reported, e.g. an equivocal or not tested result see SATISFACTORY PERFORMANCE Satisfactory performance is making ten sample reports in agreement with the consensus HLA-B27 status in a calendar year Laboratories with unsatisfactory performance will receive written notification of their status and will be expected to reply to UK NEQAS for H&I detailing their corrective actions For UK laboratories, unsatisfactory performance will be reported to UK NQAAP for Immunology For UK laboratories, failure to reply or replies deemed unsatisfactory are likely to be further actioned by UK NQAAP for Immunology. 7.5 INFORMATION/ANALYSIS PROVIDED FOR PARTICIPANTS The HLA type of the donor samples Summary sheets of all reported HLA-B27 results Information on methodology Performance information on the current and previous year s samples for all laboratories. 9

11 SCHEME 2A 8. SCHEME 2A - CYTOTOXIC CROSSMATCHING PURPOSE To assess participants ability to correctly determine cell/serum cytotoxic crossmatch status. The Steering Committee acknowledges that this crossmatching scheme will only partially emulate current crossmatching practice. NEW FOR 2014 Participants are invited to submit results obtained after dithiothreitol treatment. 8.1 SAMPLES A total of ten blood samples and forty serum samples will be sent each year as five distributions. Each distribution will comprise of two blood samples and their two corresponding sets of four selected sera (approximately 150µl each). A serum set may include test serum replicates Each set of four sera must be tested against its corresponding blood sample Participants may test peripheral blood lymphocytes PBL/T-cells and/or B-cells, according to their local practice Participants may additionally submit results for PBL/T-cells and B-cells after dithiothreitol treatment. 8.2 REPORTING At registration participants may opt for PBL/T-cell and/or B-cell crossmatch assessment Test results should be reported as positive or negative using established local criteria Tests reported as weakly positive will be interpreted as positive for assessment purposes Tests reported as equivocal will not be assessed Participants are requested to report reaction strength, using their own scoring system, to enable comparison between laboratories Participants must only use the reporting forms provided Participants are required to make their report within 10 days. 8.3 ASSESSMENT The crossmatch status of each sample is determined by at least 75% of laboratories agreeing on the positivity or negativity of each test Crossmatching tests failing to reach the 75% consensus level will not be assessed PBL/T-cell and B-cell results are considered independently Assessment Procedure A result in agreement with the consensus findings A result not in agreement with the consensus findings Acceptable Unacceptable Each sample not reported, e.g. an equivocal, void or not tested result see PBL/T-cell and B-cell results obtained after the dithiothreitol treatment are considered independently. 10

12 8.3.6 The results obtained after the dithiothreitol treatment will not be subject to the performance criteria in 2014 and will be formally assessed in 2015 SCHEME 2A 8.4 SATISFACTORY PERFORMANCE Satisfactory performance is making 85% of reports on all sera in agreement with the consensus findings in a calendar year Laboratories with unsatisfactory performance will receive written notification of their status and will be expected to reply to UK NEQAS for H&I detailing their corrective actions For UK laboratories, unsatisfactory performance will be reported to UK NQAAP for Immunology For UK laboratories, failure to reply or replies deemed unsatisfactory are likely to be further actioned by UK NQAAP for Immunology. 8.5 INFORMATION/ANALYSIS PROVIDED FOR PARTICIPANTS The HLA phenotype of the donor and specificities of the sera A summary sheet of all crossmatching results Performance information on the current and previous year s samples Performance information (non-assessed) on hidden replicate sera. 11

13 SCHEME 2B 9. SCHEME 2B - CROSSMATCHING BY FLOW CYTOMETRY PURPOSE To assess participants ability to correctly determine cell/serum flow cytometry crossmatch status. The Steering Committee acknowledges that this crossmatching scheme will only partially emulate current crossmatching practice. 9.1 SAMPLES A total of ten blood samples and forty serum samples will be sent each year as five distributions. Each distribution will comprise of two blood samples and their two corresponding sets of four selected sera (approximately 300µl each). A serum set may include test serum replicates Each set of four sera must be tested against its corresponding blood sample for IgG antibody binding. 9.2 REPORTING At registration participants may opt for T-cell and/or B-cell crossmatch assessment Test results should be reported as positive or negative compared to the local negative control Tests reported as weakly positive will be interpreted as positive for assessment purposes Tests reported as equivocal will not be assessed Participants must only use the reporting forms provided Participants are required to make their report within 10 days. 9.3 ASSESSMENT The crossmatch status of each sample is determined by at least 75% of laboratories agreeing on the positivity or negativity of each test Crossmatching tests failing to reach the 75% consensus level will not be assessed T-cell and B-cell results will be considered independently Assessment Procedure A result in agreement with the consensus findings A result not in agreement with the consensus findings Acceptable Unacceptable Each sample not reported, e.g. an equivocal, void or not tested result see SATISFACTORY PERFORMANCE Satisfactory performance is making 85% of reports on all sera in agreement with the consensus findings in a calendar year Laboratories with unsatisfactory performance will receive written notification of their status and will be expected to reply to UK NEQAS for H&I detailing their corrective actions For UK laboratories, unsatisfactory performance will be reported to UK NQAAP for Immunology For UK laboratories, failure to reply or replies deemed unsatisfactory are likely to be further actioned by UK NQAAP for Immunology. 12

14 SCHEME 2B 9.5 INFORMATION/ANALYSIS PROVIDED FOR PARTICIPANTS The HLA phenotype of the donor and the specificity of the sera A summary sheet of all reported crossmatching results Performance information on the current and previous year s samples for all laboratories Performance information (non-assessed) on hidden replicate sera A methodology questionnaire will be sent out at the end of each year. The information obtained will be summarised and distributed to participants. 13

15 SCHEME SCHEME 3 - HLA ANTIBODY SPECIFICITY ANALYSIS PURPOSE To assess participants ability to correctly determine HLA antibody specificities SAMPLES A total of ten serum samples will be sent each year as two distributions of five samples. Volumes of approximately 1.5ml of serum will be distributed REPORTING At registration participants may opt for Class I only or Class I and Class II antibody assessment Only those specificities detailed on the reporting forms will be assessed, e.g. reports of anti-a9, Bw4 and Bw6 will not be assessed Participants may report other antibody findings, e.g. DPB, DQA and MICA. These specificities will not be assessed in Participants must only use the reporting forms provided Participants are required to report antibody specificities within a period of 10 weeks ASSESSMENT Class I and Class II IgG specificities will be assessed independently Consensus presence of a specificity is determined by at least 75% of laboratories agreeing the specificity Consensus absence of a specificity is determined by at least 95% of laboratories agreeing the absence of the specificity Results failing to reach the consensus levels above will not be assessed Assessment Procedure Assigning a consensus specificity Missing a consensus specificity Assigning a specificity where the consensus is negative Acceptable Unacceptable Unacceptable Each sample report of a not tested result see SATISFACTORY PERFORMANCE Satisfactory performance is testing ten serum samples and getting at least 75% of specificities in agreement with the consensus findings in a calendar year Laboratories with unsatisfactory performance will receive written notification of their status and will be expected to reply to UK NEQAS for H&I detailing their corrective actions For UK laboratories, unsatisfactory performance will be reported to UK NQAAP for Immunology For UK laboratories, failure to reply or replies deemed unsatisfactory are likely to be further actioned by UK NQAAP for Immunology. 14

16 SCHEME INFORMATION/ANALYSIS PROVIDED FOR PARTICIPANTS Where known, the HLA phenotypes of the serum donors Summary sheets of all reported antibody findings Information on methodology Performance information on the current and previous year s samples for all laboratories. 15

17 SCHEME 4A1 11. SCHEME 4A1 DNA HLA TYPING AT 1 ST FIELD RESOLUTION PURPOSE To assess participants ability to correctly determine HLA alleles at the 1 st field level (formerly known as low resolution or 2-digit typing) SAMPLES A total of ten blood samples will be sent each year as two distributions of five blood samples REPORTING Participants may register for any of the following: HLA-A, B, C, DRB1, DRB3, DRB4, DRB5, DQA1, DQB1, and DPA1 for 1 st field assessment, or presence of assessment for DRB3, DRB4 and, DRB5. DPA1 results will be assessed if sufficient results are received Participants must only use the reporting forms provided Participants are required to return results within 4 weeks ASSESSMENT Participating laboratories will be assessed on the loci they designate at registration The consensus full HLA genotype is determined by at least 75% of laboratories agreeing each allele Alleles failing to reach the 75% consensus level will not be assessed A "blank" forms part of the assessment if at least 75% of laboratories report a single allele at a locus Participants will only be assessed on those alleles that appear in the most recently published full HLA Nomenclature Report Assessment Procedure Each full HLA genotype in agreement with the consensus 1st field type Each full HLA genotype not in agreement with the consensus 1st field type Acceptable Unacceptable Each sample report of a not tested result see SATISFACTORY PERFORMANCE Satisfactory performance is obtaining nine or more full HLA genotypes in agreement with the consensus genotypes in a calendar year Laboratories with unsatisfactory performance will receive written notification of their status and will be expected to reply to UK NEQAS for H&I detailing their corrective actions For UK laboratories, unsatisfactory performance will be reported to UK NQAAP for Immunology For UK laboratories, failure to reply or replies deemed unsatisfactory are likely to be further actioned by UK NQAAP for Immunology INFORMATION/ANALYSIS PROVIDED FOR PARTICIPANTS Summary sheets of all reported alleles Information on methodology. 16

18 Performance information on the current and previous year s samples for all laboratories. 17

19 SCHEME 4A2 12. SCHEME 4A2 DNA HLA TYPING TO 2 ND FIELD RESOLUTION PURPOSE To assess participants ability to correctly determine HLA alleles to the 2 nd field level (formerly known as high resolution or 4-digit typing) SAMPLES A total of ten blood samples will be sent each year as two distributions of five blood samples REPORTING For typing to the 2 nd field, HLA alleles should be assigned on the basis of differences in exons 2 and 3 for class I and exon 2 for class II, as a minimum requirement Participants registered for 2 nd field assessment should define all ambiguities that encompass a null allele wherever the polymorphism is located (see statement on EFI Standard D1.4 page 4) Participants may register for any of the following: HLA-A, B, C, DRB1, DRB3, DRB4, DRB5, DQA1, DQB1, DPA1 and DPB1, for 2 nd field assessment Participants must only use the reporting forms provided Participants are required to return results within 6 weeks ASSESSMENT Participating laboratories will be assessed on the loci they designate at registration The consensus full HLA genotype is determined by at least 75% of laboratories agreeing each allele Alleles failing to reach the 75% consensus level will not be assessed A "blank" forms part of the assessment if at least 75% of laboratories report a single allele at a locus Participants will only be assessed on those alleles that appear in the most recently published full HLA Nomenclature Report Assessment Procedure Each full HLA genotype in agreement with the consensus 1st & 2nd field type Each full HLA genotype not in agreement with the consensus 1st & 2nd field type Acceptable Unacceptable Each sample report of a not tested result see SATISFACTORY PERFORMANCE Satisfactory performance is obtaining nine or more full HLA genotypes in agreement with the consensus genotypes in a calendar year Laboratories with unsatisfactory performance will receive written notification of their status and will be expected to reply to UK NEQAS for H&I detailing their corrective actions For UK laboratories, unsatisfactory performance will be reported to UK NQAAP for Immunology For UK laboratories, failure to reply or replies deemed unsatisfactory are likely to be further actioned by UK NQAAP for Immunology. 18

20 SCHEME 4A INFORMATION/ANALYSIS PROVIDED FOR PARTICIPANTS Summary sheets of all reported alleles Information on methodology Performance information on the current and previous year s samples for all laboratories Sheets detailing i) the sequences that are identical over exons 2 and 3 for class I and exon 2 for class II and ii) ambiguous typing combinations (heterozygous positions identified by sequencing) defined over these exons Participants reporting strings of alleles containing null alleles will be notified. 19

21 SCHEME 4B 13. SCHEME 4B - ABO GROUPING BY DNA-BASED METHODS PURPOSE To assess participants ability to correctly determine ABO blood groups by DNA-based methods SAMPLES Laboratories performing ABO grouping by DNA-based methods, e.g. PCR-SSP, are invited to ABO group all samples from Scheme 4A REPORTING The ABO alleles detected should be reported as fully as possible, using the appropriate nomenclature Participants must only use the reporting forms provided Participants are required to return results within 4 weeks ASSESSMENT The consensus ABO genotype of each sample is determined by at least 75% of the participating laboratories agreeing the genotype Genotypes failing to reach the 75% consensus level will not be assessed Assessment Procedure Each sample report in agreement with the consensus ABO genotype Each sample report not in agreement with the consensus ABO genotype Acceptable Unacceptable Each sample report of not tested result see SATISFACTORY PERFORMANCE Satisfactory performance is making ten sample reports in agreement with the consensus ABO genotype in a calendar year Laboratories with unsatisfactory performance will receive written notification of their status and will be expected to reply to UK NEQAS for H&I detailing their corrective actions For UK laboratories, unsatisfactory performance will be reported to UK NQAAP for Immunology For UK laboratories, failure to reply or replies deemed unsatisfactory are likely to be further actioned by UK NQAAP for Immunology INFORMATION/ANALYSIS PROVIDED FOR PARTICIPANTS Summary sheets of all reported ABO genotypes Information on methodology Performance information on the current and previous year s samples for all laboratories. NOTE UK NEQAS for H&I has offered this Scheme since 2002; it is not intended to replace participation in the UK NEQAS for Blood Transfusion Laboratory Practice Scheme for laboratories using serological methods for ABO grouping. 20

22 SCHEME 5A 14. SCHEME 5A HFE TYPING PURPOSE To assess participants ability to correctly determine HFE mutations SAMPLES A total of ten blood samples will be sent each year as two distributions of five blood samples REPORTING All samples must be tested for the H63D (Hist63Asp) mutation and the C282Y (Cys282Tyr) mutation of the HFE gene Participants may register to have their Ser65Cys results assessed Please report using the single letter amino acid code, i.e. H and/or D for codon 63, C and/or Y for codon 282 and S and/or C for codon Participants are requested to report any other HFE gene mutations that they detect for participant information purposes Participants must only use the reporting forms provided Participants are required to return results within 4 weeks ASSESSMENT The consensus HFE mutations for assessment are determined by at least 75% of laboratories agreeing the combination of H63D and C282Y mutations and at least 75% of laboratories agreeing the S65C mutation Mutations failing to reach the 75% consensus level will not be assessed Assessment Procedure Each sample report in agreement with the consensus H63D and C282Y and S65C status (if applicable) Each sample report not in agreement with the consensus H63D and C282Y and S65C status (if applicable) Acceptable Unacceptable Each sample not reported, e.g. an equivocal, partial or not tested result see SATISFACTORY PERFORMANCE Satisfactory performance is making ten sample reports in full agreement with the consensus H63D and C282Y and S65C status (if applicable) in a calendar year Laboratories with unsatisfactory performance will receive written notification of their status and will be expected to reply to UK NEQAS for H&I detailing their corrective actions For UK laboratories, unsatisfactory performance will be reported to UK NQAAP for Immunology For UK laboratories, failure to reply or replies deemed unsatisfactory are likely to be further actioned by UK NQAAP for Immunology. 21

23 SCHEME 5A 14.5 INFORMATION/ANALYSIS PROVIDED FOR PARTICIPANTS A summary sheet of all reported HFE codon 63, 65 and 282 assignments Information on methodology Performance information on the current and previous year s samples for all laboratories. 22

24 SCHEME 5B SCHEME 5B - INTERPRETATIVE: HFE GENOTYPE AND HEREDITARY HAEMOCHROMATOSIS PURPOSE To assess participants ability to make an accurate, clear and concise clinical report, appropriate for the range of clinical staff involved in a patient s care and treatment, given HFE genotype and other relevant clinical information CLINICAL SCENARIOS HFE genotype will be provided, together with various pieces of clinical information, on two patients twice a year REPORTING Participants are expected to make a report on each of the patient scenarios Reports must be written in English and must be identical in format to that used for routine clinical reporting Participants are required to return their reports within 4 weeks ASSESSMENT For each of the patient scenarios several interpretative criteria expected to be covered by the report, will be identified and agreed by the expert assessors Scoring Procedure Penalty points Each feature in agreement with an identified criterion 0 A principal agreed interpretative criterion not covered by the report up to 3 Other agreed interpretative criterion not covered by the report 1 Significant erroneous patient identifiers or other information errors in the report 1 (each error) NOTE: The Expert Assessors determine the principal and other interpretative criteria Assessment Procedure Each scenario where 50% or less of the possible penalty points is allocated Each scenario where more than 50% of the possible penalty points is allocated Acceptable Unacceptable 15.4 SATISFACTORY PERFORMANCE Satisfactory performance is obtaining 4 Acceptable classifications in a year. NOTE: Unsatisfactory Performance Notifications (see Prospectus, page 6) are NOT sent to participants of Interpretative Schemes INFORMATION / ANALYSIS PROVIDED TO PARTICIPANTS Each laboratory will receive an itemisation of interpretative points applicable to each clinical scenario indicating how their report coincided with the criteria identified A summary of all participants scores General comments from the Expert Assessors on each scenario. 23

25 SCHEME SCHEME 6 HLA ANTIBODY DETECTION PURPOSE To assess participants ability to correctly determine the presence of HLA antibodies SAMPLES A total of twenty serum samples will be sent each year as two distributions of ten serum samples. Volumes of approximately 1.5ml of serum will be distributed REPORTING At registration participants may opt for Class I only or Class I and Class II antibody assessment Participants are required to report the presence or absence of HLA Class I or HLA-Class I and Class II antibody Participants must only use the reporting forms provided Participants are required to return their results within 10 weeks ASSESSMENT Participating laboratories will be assessed on the antibody class or classes they designate at registration The consensus findings for Class I and Class II specificities are determined separately Consensus positivity or negativity of each sample is determined by at least 75% of laboratories agreeing on the presence or absence of Class I or Class II antibody Samples failing to reach the 75% consensus level will not be assessed Assessment Procedure Each report in agreement with the consensus presence/absence of Class I or both Class I and Class II antibody (see and ) Each report not in agreement with the consensus presence/absence of Class I or both Class I and Class II antibody (see and ) Acceptable Unacceptable Each sample not reported, e.g. an equivocal or not tested result see The reports of participants registered for Class I and II assessment must be in agreement with the consensus Class I and Class II findings on a serum to achieve an Acceptable classification SATISFACTORY PERFORMANCE Satisfactory performance is making 80% of reports on all sera in agreement with the consensus Class I or both the consensus Class I and Class II antibody (see and above) findings in a calendar year Laboratories with unsatisfactory performance will receive written notification of their status and will be expected to reply to UK NEQAS for H&I detailing their corrective actions For UK laboratories, unsatisfactory performance will be reported to UK NQAAP for Immunology For UK laboratories, failure to reply or replies deemed unsatisfactory are likely to be further actioned by UK NQAAP for Immunology. 24

26 SCHEME INFORMATION/ANALYSIS PROVIDED FOR PARTICIPANTS Summary sheets of all reported antibody findings Information on methodology Performance information on the current and previous year s samples for all laboratories. 25

27 SCHEME SCHEME 7 HLA-B*57:01 TYPING FOR DRUG HYPERSENSITIVITY PURPOSE To assess participants ability to correctly determine HLA-B*57:01 status SAMPLES A total of ten blood samples will be sent each year as two distributions of five blood samples REPORTING Participants are required to report on HLA-B*57:01 positivity or negativity Participants may report a B*57 - non-b*57:01 - allele for information purposes Participants that routinely test to the 1 st field level only for clinical purposes will be expected to outline their reporting procedures for consideration by the UK NEQAS for H&I Steering Committee Participants must only use the reporting forms provided Participants are required to return results within 10 days ASSESSMENT The consensus HLA-B*57:01 status of each sample is determined by at least 75% of laboratories agreeing on the presence or absence of HLA-B*57: Results failing to reach the 75% consensus level will not be assessed Assessment Procedure Each report in agreement with the consensus HLA-B*57:01 status Each report not in agreement with the consensus HLA-B*57:01 status Acceptable Unacceptable Each sample not reported, e.g. an equivocal or not tested result see SATISFACTORY PERFORMANCE Satisfactory performance is making ten sample reports in agreement with the consensus HLA-B*57:01 status in a calendar year Laboratories with unsatisfactory performance will receive written notification of their status and will be expected to reply to UK NEQAS for H&I detailing their corrective actions For UK laboratories, unsatisfactory performance will be reported to UK NQAAP for Immunology For UK laboratories, failure to reply or replies deemed unsatisfactory are likely to be further actioned by UK NQAAP for Immunology INFORMATION/ANALYSIS PROVIDED FOR PARTICIPANTS Summary sheets of all reported HLA-B*57 results Information on methodology Performance information on the current and previous year s samples for all laboratories. 26

28 SCHEME SCHEME 8 HLA AND DISEASE TYPING FOR HLA-DR/DQ/DP ONLY PURPOSE To assess participants ability to correctly determine HLA-DR/DQ/DP allele families/alleles SAMPLES A total of ten DNA preparations will be sent each year as two distributions of five DNA preparations These samples have been previously tested in Scheme 4A REPORTING Participants are required to report their HLA-DR and/or HLA-DQ and/or HLA-DP findings Participants must only use the reporting forms provided Participants are required to return results within 6 weeks ASSESSMENT Participating laboratories will be assessed on the loci typed for each sample Assessment will be at the 1 st or 2 nd field level appropriate to the individual participant s results for each locus and for each sample. DRB3/4/5 results at the presence or absence level will also be accepted The consensus genotype for each locus is previously determined by at least 75% of laboratories agreeing each allele or blank in Scheme 4A Reports of groups of alleles will be assessed at the allele family level Participants will only be assessed on those alleles that appear in the most recently published full HLA Nomenclature Report Assessment Procedure Each HLA locus in agreement with the 1 st or 2nd field consensus type (see ) Acceptable Each HLA locus not in agreement with 1 st or 2nd field the consensus type (see ) Unacceptable Each sample not reported see SATISFACTORY PERFORMANCE Satisfactory performance is obtaining nine or more full HLA genotypes in agreement with the consensus genotypes in a calendar year Laboratories with unsatisfactory performance will receive written notification of their status and will be expected to reply to UK NEQAS for H&I detailing their corrective actions For UK laboratories, unsatisfactory performance will be reported to UK NQAAP for Immunology For UK laboratories, failure to reply or replies deemed unsatisfactory are likely to be further actioned by UK NQAAP for Immunology INFORMATION/ANALYSIS PROVIDED FOR PARTICIPANTS Summary sheets of all reported HLA-DR, DQ and DP results and the original HLA type derived from Scheme 4A2 testing Information on methodology Performance information on the current year s samples for all laboratories. 27

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