UK National External Quality Assessment Research Articles

Evaluation of cryoprotein investigation using a digital external quality assurance scheme.

Background: Robust preanalytical and analytical processes are critical for the detection of cryoproteins. There is significant variation in practice in the detection, analysis and reporting. Results: A survey in 2018 of 137 laboratories participating in the UK National External Quality Assessment Service (UK NEQAS) (6) quality control program showed significant variation in the laboratory processes which highlighted the need for standardisation of the detection, analysis and reporting of cryoglobulins.Conclusion: The first available EQA scheme aiming to harmonise practice for cryoprotein testing has been developed by UK NEQAS and laboratories should participate in an appropriate EQA scheme to fulfil requirements for ISO accreditation.

PB1112 UK National External Quality Assessment Scheme for Blood Coagulation (UK NEQAS BC) Emicizumab Programme, Survey July 2022 Results for Modified One-Stage Assay and Chromogenic Assay

PB1112 UK National External Quality Assessment Scheme for Blood Coagulation (UK NEQAS BC) Emicizumab Programme, Survey July 2022 Results for Modified One-Stage Assay and Chromogenic Assay

PB0616 UK National External Quality Assessment Scheme for Blood Coagulation (UK NEQAS BC) Evaluation of the Point of Care Fibrinogen Device

PB0616 UK National External Quality Assessment Scheme for Blood Coagulation (UK NEQAS BC) Evaluation of the Point of Care Fibrinogen Device

Internal Quality Control in Hemostasis Assays.

Internal quality control (IQC) for routine and specialist hemostasis testing represents a mandatory requirement for assays offered by clinical laboratories under International Organization for Standardization, Code of Federal Regulations, and Clinical and Laboratory Standards Institute standards. The underlying principle is that regular IQC audits the analytical performance of automated, semiautomated, and manual methods. This review investigates IQC practices, including benefits, limitations, frequency per time period or batch, sources of material used, primary supplier, third party or in-house, plus troubleshooting when IQC falls outside acceptance criteria. To assess IQC practice, the UK National External Quality Assessment Scheme (NEQAS) Blood Coagulation distributed a questionnaire to 1,200 participants enrolled in our scheme that collected details of the local practices for IQC testing. We received returns from 127 centers that described their local practices for the frequency of IQC, the type of IQC material employed, acceptance criteria for IQC data, and troubleshooting protocols for IQC failures. The data collected as part of an NEQAS BC questionnaire confirmed that all the participants returning answers to the questionnaire meet the standards for regular IQC testing for the hemostasis assays they perform.

The history and evolution of HLA typing external proficiency testing schemes in UK NEQAS for H&I.

The UK National External Quality Assessment Service (NEQAS) provide an external proficiency testing (EPT) service for clinical laboratories. UK NEQAS for Histocompatibility and Immunogenetics (H&I) has been providing EPT schemes for over 45years and has grown during this time to provide 19 EPT schemes. Accurate human leucocyte antigen (HLA) typing is critical to support safe clinical services, including transplantation, therefore high quality, relevant EPT schemes are required as part of a laboratory's quality assurance. This article reviews the development of the HLA typing EPT schemes, from the first HLA phenotyping scheme in 1975, via the first HLA genotyping scheme in 1992, through to the introduction in 2017 of HLA third field assessment results from next-generation sequencing technology. In addition, the introduction of EPT schemes to cover HLA associated diseases and pharmacogenetic reactions, including HLA-B27, HLA*B*57:01 and HLA-DQ for coeliac disease are discussed. The accuracy of laboratory EPT results for HLA phenotyping are >96% (2018-2022), HLA genotyping >99% (2020-2022), HLA-B27 testing >99% (2018-2022) and B*57:01 testing >99% (2017-2022). However, for HLA genotyping for coeliac disease 22%-46% of laboratories made errors in 2020-2022. On investigation, the high rate of unsatisfactory performance was attributed to laboratories lacking specific knowledge to interpret HLA genotyping results and accurately report HLA types for coeliac disease. A misleading commercial kit insert was also identified. The assessment of scheme results has uncovered several issues which have been addressed with the intention of educating participants and improving clinical services. The UK NEQAS for H&I EPT schemes have evolved over the past four decades to reflect changes in HLA typing technology, laboratory clinical practice and to cover post-analytical interpretative elements of HLA typing.

DS23 Efficiency and performance of slide stainers during Mohs frozen section processing

Abstract Mohs micrographic surgery (MMS) relies on good-quality, well-stained frozen sections for effective histological analysis. Slides should also be produced as quickly as possible. Automation of complementary devices in the laboratory ensures standardization and efficiency and improves the overall quality of performance. Our study evaluated the efficiency and performance of the Autostainer-XL (Leica) vs. the Linistat (Thermo Scientific) slide stainer within a busy MMS unit. The Autostainer-XL rapidly dries, loads and unloads slides automatically allowing the Mohs-trained biomedical scientist (BMS) to work in parallel on cryotomy. The Autostainer-XL stains slides with haematoxylin and eosin within 10 min and 50 s. In comparison, the Linistat was much slower and required the Mohs-trained BMS to fix the slides manually on a hotplate, place them in industrial methylated spirit, wash them in water and load individual slides into the Linistat, taking 16 min and 45 s to complete the staining process. Two blinded assessors retrospectively reviewed the same set of 50 Linistat-stained and 50 Autostainer-XL stained frozen section slides for consistency in staining. The scoring criteria were based on the modified UK National External Quality Assessment Service (NEQAS) Cellular Pathology Technique Mohs scheme assessment criteria. Each assessor allocated scores between 1 and 5 based on the scoring criteria. The mean quality scores were significantly higher for the Autostainer-XL compared with the Linistat (P < 0.05). In our MMS unit, up to 12 patients undergo MMS daily with an average of seven slides required per patient at the first stage, leading to a high processing burden within the Mohs laboratory, particularly at the first stage. The Autostainer-XL increased productivity, with the ability to load 30 slides per rack, while the Linistat was only able to load six slides per rack. In practice, within our MMS unit, the Autostainer-XL reduced the time taken to produce all slides at the first layer by more than 1 h when compared with the Linistat slide stainer. When surveyed, 100% (n = 8/8) of Mohs-trained BMS in the laboratory preferred the Autostainer-XL than the Linistat, with comments suggesting that it was easy to use, quick, precise and an important device that can withstand the demands of a busy Mohs laboratory. The Autostainer-XL offers improved efficiency and performance, with reduced staining times, an ability to process a larger number of slides and fully customisable automation when compared with the Linistat (Thermo Scientific) slide stainer.

Results from the second WHO external quality assessment for the molecular detection of respiratory syncytial virus, 2019-2020.

External quality assessments (EQAs) for the molecular detection of human respiratory syncytial virus (RSV) are necessary to ensure the standardisation of reliable results. The Phase II, 2019-2020 World Health Organization (WHO) RSV EQA included 28 laboratories in 26 countries. The EQA panel evaluated performance in the molecular detection and subtyping of RSV-A and RSV-B. This manuscript describes the preparation, distribution, and analysis of the 2019-2020 WHO RSV EQA. Panel isolates underwent whole genome sequencing and in silico primer matching. The final panel included nine contemporary, one historical virus and two negative controls. The EQA panel was manufactured and distributed by the UK National External Quality Assessment Service (UK NEQAS). National laboratories used WHO reference assays developed by the United States Centers for Disease Control and Prevention, an RSV subtyping assay developed by the Victorian Infectious Diseases Reference Laboratory (Australia), or other in-house or commercial assays already in use at their laboratories. An in silico analysis of isolates showed a good match to assay primer/probes. The panel was distributed to 28 laboratories. Isolates were correctly identified in 98% of samples for detection and 99.6% for subtyping. The WHO RSV EQA 2019-2020 showed that laboratories performed at high standards. Updating the composition of RSV molecular EQAs with contemporary strains to ensure representation of circulating strains, and ensuring primer matching with EQA panel viruses, is advantageous in assessing diagnostic competencies of laboratories. Ongoing EQAs are recommended because of continued evolution of mismatches between current circulating strains and existing primer sets.

UK NEQAS Haematology pilot scheme: Reticulocyte haemoglobin content quantitation.

Established parameters for the investigation of suspected iron deficiency have recognized limitations that affect their sensitivity and specificity. Reticulocyte haemoglobin content (RHC) is an early biomarker of iron deficiency or restriction, which also demonstrates response to iron therapy. RHC parameters are offered on all major automated haematology analysers. There is no external quality assessment (EQA) for RHC in the UK, restricting its wider clinical application. The UK National External Quality Assessment Scheme (UKNEQAS) for Haematology has completed a pilot study on the use of stabilized blood for RHC EQA. Blood was obtained from two JAK2 V617F mutation-positive, polycythaemia vera patients undergoing regular venesection, and from a healthy volunteer donor. Aliquots of each donation were distributed to users of Abbott, Horiba, Siemens and Sysmex automated analysers for RHC analysis on days 1, 3, 5 and 7. Results were received from 20 laboratories using four different platforms. The daily mean and standard deviation (SD) were calculated for each aliquot, by day of analysis and platform. With RHC there was no significant within-platform difference (p> 0.5) between testing days, although there were statistically significant differences (p< 0.001) between different platforms. The greatest difference was seen between Abbott MCHr and Horiba RhCc (-6.14 ± 1.25 pg). Despite inter-platform differences, it was possible to define iron deficient, borderline, and normal cut-offs to classify 95% of results correctly. The results demonstrate that it is possible to provide RHC EQA material using UKNEQAS standard procedures. The results are clinically relevant but indicate a requirement for inter-user comparison.

External quality assessment for one-stage and chromogenic factor IX assays in samples containing Alprolix (rFIXFc) or Idelvion (rIX-FP) and evidence that UK National External Quality Assessment Scheme for blood coagulation samples are commutable with patient samples.

There may be clinically relevant differences between results of different FIX assays in samples containing extended half life FIX concentrates requiring regular surveillance of assay results through proficiency testing exercises. Control materials used in proficiency testing must be commutable, that is have the same inter-assay properties as those demonstrated by authentic clinical samples when measured by different analytical methods. We assessed relationships between results with different FIX assays and commutability of UK National External Quality Assessment Scheme (NEQAS) materials containing rIX-FP (Idelvion) or rFIXFc (Alprolix) by comparing results obtained using widely used one-stage and chromogenic assays during a proficiency testing exercise with results obtained when analysing a series of individual patient samples using the same assay systems. NEQAS samples prepared by addition of either Idelvion or Alprolix to FIX deficient plasma were sent to 76haemophilia centres in Europe. A total of 18 Idelvion and 22 Alprolix patient samples were assayed in a single centre. Two chromogenic and two one-stage assays were compared. The pattern of results obtained for NEQAS samples and patient samples was similar. In all cases, the NEQAS sample data point was within the scatter of patient sample data in plots of patient sample results showing one-stage assay results using Synthasil or Actin FS plotted against chromogenic assay results with Biophen or Rox chromogenic FIX kits. This indicates that the NEQAS samples containing Idelvion or Alprolix were commutable and therefore suitable for use in proficiency testing exercises.

Anti‐PF4 testing for vaccine‐induced immune thrombocytopenia and thrombosis and heparin induced thrombocytopenia: Results from a UK National External Quality Assessment Scheme exercise April 2021

BackgroundVaccine‐induced immune thrombocytopenia and thrombosis (VITT) following the administration of the AstraZeneca (AZ) ChAdOx1 nCOV‐19 vaccine has recently been reported. The associated clinical and laboratory features have included thrombosis at unusual sites, thrombocytopenia, and raised D‐dimers with positivity for IgG anti‐platelet factor 4 (PF4) antibodies. ObjectivesA UK National External Quality Control Assessment Scheme external quality control exercise was carried out by distributing liquid and lyophilized samples from a subject with VITT, a pool of samples from subjects with classical heparin‐induced thrombocytopenia (HIT), and a non‐VITT/non‐HIT case to 85 centers performing HIT testing. MethodsParticipating centers employed their locally validated testing methods for HIT assays. ResultsThe lyophilized and liquid samples were found to be commutable for the ELISA assays used in the detection of anti‐PF4 antibodies. The Aeskulisa, Stago, Hyphen, and LIFECODES anti‐PF4 ELISA assays successfully detected the VITT antibody, whereas the Acustar HIT, Werfen LIA, and the Stago STIC assays did not. ConclusionIt is important that clinical and laboratory teams are aware of the limitations of some anti‐PF4 assays when using them to aid diagnosis of VITT syndrome.

The Use and Effectiveness of an Online Diagnostic Support System for Blood Film Interpretation: Comparative Observational Study.

BackgroundThe recognition and interpretation of abnormal blood cell morphology is often the first step in diagnosing underlying serious systemic illness or leukemia. Supporting the staff who interpret blood film morphology is therefore essential for a safe laboratory service. This paper describes an open-access, web-based decision support tool, developed by the authors to support morphological diagnosis, arising from earlier studies identifying mechanisms of error in blood film reporting. The effectiveness of this intervention was assessed using the unique resource offered by the online digital morphology Continuing Professional Development scheme (DM scheme) offered by the UK National External Quality Assessment Service for Haematology, with more than 3000 registered users. This allowed the effectiveness of decision support to be tested within a defined user group, each of whom viewed and interpreted the morphology of identical digital blood films.ObjectiveThe primary objective of the study was to test the effectiveness of the decision support system in supporting users to identify and interpret abnormal morphological features. The secondary objective was to determine the pattern and frequency of use of the system for different case types, and to determine how users perceived the support in terms of their confidence in decision-making.MethodsThis was a comparative study of identical blood films evaluated either with or without decision support. Selected earlier cases from the DM scheme were rereleased as new cases but with decision support made available; this allowed a comparison of data sets for identical cases with or without decision support. To address the primary objectives, the study used quantitative evaluation and statistical comparisons of the identification and interpretation of morphological features between the two different case releases. To address the secondary objective, the use of decision support was assessed using web analytical tools, while a questionnaire was used to assess user perceptions of the system.ResultsCases evaluated with the aid of decision support had significantly improved accuracy of identification for relevant morphological features (mean improvement 9.8%) and the interpretation of those features (mean improvement 11%). The improvement was particularly significant for cases with higher complexity or for rarer diagnoses. Analysis of website usage demonstrated a high frequency of access for web pages relevant to each case (mean 9298 for each case, range 2661-24,276). Users reported that the decision support website increased their confidence for feature identification (4.8/5) and interpretation (4.3/5), both within the context of training (4.6/5) and also in their wider laboratory practice (4.4/5).ConclusionsThe findings of this study demonstrate that directed online decision support for blood morphology evaluation improves accuracy and confidence in the context of educational evaluation of digital films, with effectiveness potentially extending to wider laboratory use.

Factor VIII/IX inhibitor testing practices in the United Kingdom: Results of a UKHCDO and UKNEQAS national survey.

Inhibitor formation is the greatest challenge facing persons with haemophilia treated with factor concentrates. The gold standard testing methodologies are the Nijmegen-Bethesda assay (NBA) for FVIII and Bethesda assay (BA) for FIX inhibitors, which are affected by pre-analytical and inter-laboratory variability. To evaluate inhibitor testing methodology and assess correlation between self-reported and actual methodology. Methodology was evaluated using a survey distributed alongside a UK National External Quality Assessment Service Blood Coagulation external quality assurance (EQA) exercise for FVIII and FIX inhibitor testing. Seventy four survey and EQA exercise responses were received (response rate 63.2%), with 50 paired survey/EQA results. 47.1% (33/70) reported using the NBA and 42.9% (30/70) the BA for FVIII inhibitor testing. Review of FVIII inhibitor assay methodology demonstrated discrepancy (self-reported to actual) in 64.3% (BA reporting) and 27.6% (NBA reporting). Pre-analytical heat treatment was used by 32.4%, most commonly 56°C for 30minutes. Assay cut-offs of 0.1-1.0BU/mL were reported. EQA samples (acquired FVIII and congenital FIX) demonstrated titres and coefficients of variation (CV) of 3.1BU/mL (0.7-15.4BU/mL; CV=43%) and 18.0BU/mL (0-117BU/mL; CV=33%), respectively. No significant assay or laboratory factors were found to explain this variance, which could have resulted in change in management for 6 patients (5 misclassified high-titre FVIII inhibitors and 1 false negative for a FIX inhibitor). Heterogeneity was seen at each stage of assay methodology. No assay-related factors were found to explain variation in inhibitor titres. Further standardization is required to improve inhibitor quantification to guide patient care.

WHO malaria nucleic acid amplification test external quality assessment scheme: results of distribution programmes one to three

BackgroundThe World Health Organization (WHO) recommends parasite-based diagnosis of malaria. In recent years, there has been surge in the use of various kinds of nucleic-acid amplification based tests (NAATs) for detection and identification of Plasmodium spp. to support clinical care in high-resource settings and clinical and epidemiological research worldwide. However, these tests are not without challenges, including lack (or limited use) of standards and lack of reproducibility, due in part to variation in protocols amongst laboratories. Therefore, there is a need for rigorous quality control, including a robust external quality assessment (EQA) scheme targeted towards malaria NAATs. To this effect, the WHO Global Malaria Programme worked with the UK National External Quality Assessment Scheme (UK NEQAS) Parasitology and with technical experts to launch a global NAAT EQA scheme in January 2017.MethodsPanels of NAAT EQA specimens containing five major species of human-infecting Plasmodium at various parasite concentrations and negative samples were created in lyophilized blood (LB) and dried blood spot (DBS) formats. Two distributions per year were sent, containing five LB and five DBS specimens. Samples were tested and validated by six expert referee laboratories prior to distribution. Between 37 and 45 laboratories participated in each distribution and submitted results using the online submission portal of UK NEQAS. Participants were scored based on their laboratory’s stated capacity to identify Plasmodium species, and individual laboratory reports were sent which included performance comparison with anonymized peers.ResultsAnalysis of the first three distributions revealed that the factors that most significantly affected performance were sample format (DBS vs LB), species and parasite density, while laboratory location and the reported methodology used (type of nucleic acid extraction, amplification, or DNA vs RNA target) did not significantly affect performance. Referee laboratories performed better than non-referee laboratories.ConclusionsGlobally, malaria NAAT assays now inform a range of clinical, epidemiological and research investigations. EQA schemes offer a way for laboratories to assess and improve their performance, which is critical to safeguarding the reliability of data and diagnoses especially in situations where various NAAT methodologies and protocols are in use.

Anti-pneumococcal antibodies (anti-PS): lack of standardisation between assays skews UK national external quality assessment services (UKNEQAS) consensus targets

Introduction: Anti-PS assays are used to assess immunodeficiency. Our in-house ELISA is calibrated against the FDA 89SF standard. Assay standards and samples are pre-adsorbed with capsular polysaccharide (CPS) and 22F. The UKNEQAS programme is dominated by the Binding Site VaccZyme anti-PCP (>90% participants). This ELISA uses a purified protein standard pre-adsorbed with CPS only.

Clotting and chromogenic factor VIII assay variability in post‐infusion and spiked samples containing full‐length recombinant FVIII or recombinant factor VIII Fc fusion protein (rFVIIIFc)

Variability in FVIII measurement is a recognized problem. There are limited data for samples containing recombinant Factor VIII Fc fusion protein (rFVIIIFc). Many studies use samples for which factor concentrate has been spiked into FVIII deficient plasma in vitro. This approach requires validation. Four samples were distributed in a UK National External Quality Assessment Scheme for Blood Coagulation (NEQAS BC) survey. One contained Advate (full-length recombinant FVIII) (rFVIII) added to FVIII deficient plasma, one was from a severe haemophilia A patient after infusion of Advate, one was prepared by addition of rFVIIIFc (marketed as Elocta/Eloctate) to FVIII deficient plasma and the fourth was collected from a severe haemophilia A patient following rFVIIIFc (Eloctate) infusion. Fifty-three haemophilia centres (UK and Scandinavia) performed one-stage FVIII assays and 27 performed chromogenic FVIII assays. One-stage assays gave significantly lower results than chromogenic assays by 7% (P<0.01) and 13%(P<0.001) for post-Advate and Advate spiked samples, and by 22% (P<0.001) and 23% (P<0.001) for post-rFVIIIFc and rFVIIIFc spiked samples. The interlaboratory variation was similar for all samples, with CVs of 12%-16% (chromogenic) and 10%-13% (one stage). The data indicate that either product can be safely monitored by one-stage or chromogenic assay. Spiked samples behaved in a similar way to post-infusion samples for both products and could be substituted for post-infusion samples for use in proficiency testing exercises (ie, samples were commutable).

Breast cancer biomarkers in clinical testing: analysis of a UK national external quality assessment scheme for immunocytochemistry and in situ hybridisation database containing results from 199 300 patients

We describe a collated data set of results from clinical testing of breast cancers carried out between 2009 and 2016 in the United Kingdom and Republic of Ireland. More than 199 000 patient biomarker data sets, together with clinicopathological parameters were collected. Our analyses focused on human epidermal growth factor receptor‐2 (HER2), oestrogen receptor (ER) and progesterone receptor (PR), with the aim of the study being to provide robust confirmatory evidence on known associations in these biomarkers and to uncover new data on previously undescribed or unconfirmed associations, thus strengthening the evidence‐base in clinical breast cancer testing. Overall, 13.1% of tumours were HER2‐positive; 10.6% in ER‐positive tumours, and 25.5% in ER‐negative tumours. Higher rates of HER2 positivity were significantly associated with patient age <56 years versus age ≥56 years, symptomatic versus screen‐detected tumours, testing of involved axillary node versus primary breast cancer, invasive ductal carcinoma (not otherwise specified) versus other histological types, higher histological grade, increasing tumour size, increasing nodal involvement, ER‐negative versus ER‐positive tumour status, PR‐negative versus PR‐positive tumour status. Where ER status was known, 82.7% of tumours were ER‐positive; 80.9% in women age <56 years, and 83.6% in those age ≥56 years (ER‐positive cut‐off ≥1.0% positive tumour cells or equivalent). Where PR status was known, 64.9% of tumours were PR‐positive; 65.8% in women age <56 years, and 64.4% in women age ≥56 years (PR‐positive cut off ≥10.0% or equivalent). These analyses of clinical test results provide contemporary benchmarking data for HER2, ER and PR positive rates.

The importance of commutability in material used for quality control purposes.

External Quality Assessment (EQA) is an important part of laboratory quality assurance. Spiking of normal plasma is sometimes employed to mimic clinical samples. It is important that spiked material gives similar results to clinical samples (ie, is commutable) to ensure appropriate conclusions can be drawn from EQA exercises. We describe here data from UK National External Quality Assessment Scheme for Blood Coagulation (NEQAS BC) exercises where spiked samples were tested alongside samples from patients to explore commutability of artificial material. Normal plasma was spiked with unfractionated heparin (UFH), low molecular weight heparin (LMWH), Dabigatran or Rivaroxaban. Factor VIII (FVIII) deficient plasma was spiked with FVIII concentrate. Spiked samples and ex vivo patient material were sent to laboratories for testing. For LMWH, good agreement was seen between results for samples from patient plasma and plasma spiked with heparin. For UFH, APTT ratios differed between spiked and patient samples for the same drug concentration, with no correlation in ranking of reagents. Precision for patient and spiked material for Rivaroxaban and Dabigatran assays was comparable. However, the pattern of results for some Dabigatran assay kits differed between spiked and patient samples. For FVIII assays, results obtained with spiked and postinfusion samples gave comparable results. Spiked material is suitable for EQA if commutability is demonstrated. Our data show commutability for plasma spiked with Rivaroxaban, LWMH and some FVIII concentrates. For some tests, notably APTT for UFH, marked differences between patient and spiked samples indicate not all tests can be evaluated using artificial samples.

Laboratory Performance on Reporting Monoclonal Gammopathy During Cerebrospinal Fluid Oligoclonal Banding Analysis from External Quality Assessment Surveys.

Cerebrospinal fluid (CSF) oligoclonal banding (OCB) analysis is a sensitive test used to mainly aid multiple sclerosis (MS) diagnosis. Monoclonal gammopathy is usually an incidental finding during CSF OCB analysis. The aim of this study was to assess laboratory performance on reporting monoclonal gammopathy pattern during CSF OCB analysis based on external quality assessment surveys. The CSF OCB surveys from the College of American Pathologists (CAP) from 2010 to 2015 were reviewed. The UK National External Quality Assessment Service (NEQAS) CSF OCB surveys from 2014 to 2017 were also reviewed. All monoclonal gammopathy patterns were confirmed by serum protein electrophoresis followed by immunofixation on a Sebia Hydrasys analyzer. There were 11 monoclonal gammopathy cases identified in the CAP OCB survey from 2010 to 2015. The average rate of CAP participants that correctly reported the pattern was 25.1% (range, 2.4%-66.7%). The most common pattern incorrectly reported was the systemic inflammation pattern, followed by the oligoclonal bands present/positive pattern. The NEQAS OCB survey from 2014 to 2017 had 4 monoclonal gammopathy cases and indicated a much higher number (average, 88.5%; range, 84.1%-90.8%) of participating laboratories to successfully detect monoclonal gammopathy. Monoclonal gammopathy is still an underrecognized pattern in the CSF OCB analysis by the CAP participating laboratories and warrants further education.

Abstract P3-08-16: ER, PR and HER2 biomarkers in UK and Irish clinical breast cancer testing: analysis of results from &amp;gt;168,000 patients

Abstract AIMS To describe and analyse &amp;gt;168,000 sets of results from clinical breast biomarker testing carried-out between 2009 and 2016 in the UK and Republic of Ireland, focussing on biological relationships. To present robust confirmatory evidence on known associations and provide new data on previously undescribed or unconfirmed ones. BACKGROUND Between Jan-2009 and Jan-2016 the UK National External Quality Assessment Scheme for Immunocytochemistry and In-situ Hybridisation (UK NEQAS ICC&amp;ISH) collected results on clinical breast cancer testing, comprehensively for human epidermal growth factor receptor-2 (HER2) and optionally for estrogen receptor (ER) and progesterone receptor (PR), from most UK and Irish testing centres. Primary objectives were to assess and, where indicated achieve improvements in testing quality. The size and scope of the data-set is unparalleled in the literature and represent a significant reference resource for the breast cancer community. METHODS UK NEQAS ICC&amp;ISH created and curated an on-line data-entry system allowing centres to systematically collect their own HER2, ER and PR results from clinical testing; for HER2, immunohistochemical (IHC) and in-situ hybridisation (ISH) data were collected; for ER and PR, IHC results could be entered according to local practise format. Tools and guidance were provided enabling centres to analyse their data for local audit and quality assurance. Clinicopathologic data was also collected, including: patient age at diagnosis, histological tumor type, tumor grade, site/stage (primary, recurrence, metastasis) and sample type (core, excision). RESULTS Data are present on 168,793 patients. 173 centres contributed ≥100 entries (96% of total). Median age was 62 years (IQR: 51-72). Tumor type was stated in 42%: 76% were invasive ductal, and 13% invasive lobular carcinoma. Grade was stated in 56%: 15% were Grade 1, 55% Grade 2 and 30% Grade 3. Site/stage was stated in 56%: 92% were primary, 2% recurrent and 6% metastatic. Sample type was stated in 70%: 75% were cores and 25%, excision. Receptor statuses were available as follows; HER2 (100%): 87% negative, 13% positive; ER (45%): 15% negative, 85% positive; PR (31%): 29% negative, 71% positive. HER2 data were available as follows; category by IHC: 91%; amplification status by ISH: 15%; HER2 gene/chromosome 17 (CEP17) ratio: 15% (86% of which were IHC 2+); HER2 gene copy number: 7%; CEP17 copy number: 7%. HER2-positive rate was 24% in ER-negative and 11% in ER-positive cases. HER2-positive status was negatively associated with increasing ER positivity (rho -0.22, P&amp;lt;0.001). 71% of HER2-positive cases were ER-positive and 29% were ER-negative. The HER2 2+ rate was 14% in ER-negative and 21% in ER-positive cases. Considering only IHC 2+ cases, median HER2 copy number was 5.4 in ER-negative and 4.4 in ER-positive disease. Comprehensive description of significant associations for all parameters will be presented. CONCLUSION The unique size and scope of this data-set has allowed confirmation of known associations for HER2, ER and PR with clinicopathologic and biological characteristics in breast cancer to very high confidence levels, and has uncovered previously undescribed relationships in both ER-positive and ER-negative disease. Citation Format: Dodson A, Parry S, Ibrahim M, Bartlett J, Dowsett M, Miller K. ER, PR and HER2 biomarkers in UK and Irish clinical breast cancer testing: analysis of results from &amp;gt;168,000 patients [abstract]. In: Proceedings of the 2017 San Antonio Breast Cancer Symposium; 2017 Dec 5-9; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2018;78(4 Suppl):Abstract nr P3-08-16.

UK NEQAS survey of allergen component testing across the United Kingdom and other European countries.

The clinical utility of molecular diagnostic approaches in allergy investigation is being recognized increasingly to play a significant role in the management of allergic patients. Determining the sensitization pattern, which is best achieved through the use of component resolved diagnostics (CRD), allows effective risk stratification, appropriate treatment and patient selection for immunotherapy. In order to assess the diagnostic service provisions for in-vitro allergy testing across Europe, a survey was carried out via the total immunoglobulin (Ig)E and specific IgE external quality assurance schemes run by UK National External Quality Assessment Service (NEQAS) Immunology, Immunochemistry and Allergy. This survey assessed allergy testing, and in particular allergen components offered by the laboratories, and found a wide variability in service provision, particularly between the United Kingdom and other European Union (EU) countries. Furthermore, there was lack of standardization for acquisition of clinical information to aid allergen (and component) selection, gating strategy, testing algorithms and clinical interpretation. Interestingly, a significant proportion of laboratories (the majority from EU) stated that they 'used' the results for peanut components for risk stratification. However, the vast majority of participants were unaware of guidelines relating to the use of allergen component testing, and agreed that further education would assist in reaching a common platform. Hence, this survey has highlighted that although CRD has been adopted into routine diagnostics across Europe, it is potentially compromised by lack of standardized protocols and guidance sources. Consequently, there is a need for local or national standards and education through External Quality Assurance services on the performance and application of CRD into allergy investigation.