bluuebunny/crossref_metadata_2025 huggingface dataset:
B-030 Performance evaluation of Siemens Atellica® IM Anti-Müllerian Hormone Assay
Authors: Shang E , Agustin H , El-Khoury J , Lee J K , Jones J , Scheidegger M , Abou-Diwan C , Rhea-McManus J , Li J
Published: October 2024

Background

Anti-Müllerian hormone (AMH), a glycoprotein dimer, is produced by ovarian granulosa cells and its levels have been shown to directly correlate to the number of eggs remaining in the ovaries (ovarian reserve). Thus, measurements of AMH are widely used as an aid in the assessment of the ovarian reserve in women presenting to fertility clinics. This study aimed to evaluate the performance of the Siemens Atellica® IM AMH assay and compare it to the Beckman Access AMH assay.

Methods

The Atellica AMH assay is a fully automated chemiluminescent monoclonal sandwich immunoassay, while the Beckman Access AMH assay is also an automated chemiluminescent monoclonal sandwich immunoassay. Precision and linearity were assessed for the Atellica AMH assay using manufacturer-provided standards. Inter-laboratory correlation studies were completed using deidentified patient samples collected from 120 reproductive-age women undergoing fertility outpatient evaluation at Yale New Haven Health. Blood was drawn on the first office visit for fertility workup. AMH values were initially performed at Yale University using the Beckman Access AMH assay. Serum samples were then frozen (-20°C) and shipped on dry ice to the laboratory at the Ohio State University where they were thawed and analyzed using the Siemens Atellica AMH assay. Linear regression was employed for data analysis. Overall percent agreement using an AMH cut-off value of 1.77 ng/mL was calculated. AMH values >1.77 ng/mL were classified as positive and values ≤1.77 were classified as negative.

Results

The Siemens Atellica AMH assay exhibited good performance across the measuring range, with intra-assay and inter-assay coefficients of variation (CVs) ranging from 1.4% to 3.2% and 2.5% to 6.0%, respectively. The assay demonstrated linearity over the four recommended ranges (0.043-24.0 ng/mL) without significant bias observed. The Access and Atellica AMH assays demonstrated good overall agreement of 97% and were well-correlated with a R2 value of 0.9721.

Conclusions

The AMH assay was validated and routinely measured in-house using the Atellica automated assay. The Siemens Atellica AMH assay performs well across its measuring range and shows good agreement with Beckman Access AMH at the AMH cut-off of 1.77 ng/mL.
B-031 A new Ready to use Bio-Rad Quality Control to Streamline Daily Workflow
Authors: Yagoot A M , Khalil H , Alharbi M , Almalayo G , Borai A
Published: October 2024

Background

A comprehensive Quality Management System starts with performing a daily Quality Control (Q.C.) process. The daily Q.C. process monitors the Instrument’s optimal performance and detects shifts and trends. However, the existing traditional daily Q.C. process is not only prone to manual errors, but also time consuming, costly and labor intensive. We evaluated a Bio-Rad InteliQTM Load and go Quality Control that comes in a barcoded tube ready to be loaded. We monitored the workflow efficiency gained in comparison to the traditional quality control material that comes in glass vials.

Methods

We evaluated the new quality control material ready to load on the instrument. This new Q.C. (Bio-Rad InteliQ Assayed Multiqual Control - 3 levels) were processed in parallel with our routine Q.C. (Bio-Rad Assayed Chemistry Lyphocheck vials - 2 levels). We performed the evaluation on Abbott Architect c8000 Chemistry analyzers in our central Laboratory (NGHA-Jeddah). The Q.C. was stored frozen. Before testing it was thawed at room temperature as per the insert instruction. Over a 5 days period, in parallel with our routine quality control, we run the new Q.C. Two times per day for 33 analytes. We monitored the processing time difference between the new and current approaches. We monitored the stability of all the analytes in the tube after Two days and again after Five days. We also monitored the cost effectiveness, labor intensiveness, probability of operator error and the overall improvement in workflow and Turn Around Time.

Results

Over the evaluation period, we observed the following: - Saving 40 minutes in average per week only for chemistry Q.C (time spent in reconstituting, mixing and aliquoting old Q.C) - 100% reduction in waste of plastic consumables such as pipette and plastic tubes. - Save an average of 1.0 ml dead volume in every 5.0 ml old Q.C. vial (20% of the volume, hence saving 20% of the total Q.C. cost). The dead volume wasted due to aliquoting step using the old Q.C. - 100% Eliminated delays caused by operator error such as wrong reconstitution, inadequate mixing and wrong Q.C. aliquoting. Not needed for the InteliQ. - High score of staff satisfaction. - Stability of the 32 analytes was excellent except that of Co2, which deteriorated by 14% after Two days as indicated in the insert package. Co2 day five result deteriorated by 34%.

Conclusions

Bio-Rad InteliQ is easy to use. It saves time and cost. When combined with Unity Q.C. data management solution it can streamline Q.C. workflow, eliminate operator errors, reduce T.A.T. and increase the overall laboratory performance. It may also work optimally with Instruments that have on-board fridge for Q.C.
B-035 Atellica CI Analyzer Sample-to-sample Carryover Performance Evaluation
Authors: Kampfrath T , Diabate D , Maxwell T , Seltmann J , Shalhoub V , Lewisch S , Snyder J
Published: October 2024

Background

Laboratories in many parts of the world are required to demonstrate/confirm the lack of sample carryover on their analyzer under their laboratory conditions using patient populations. Sample carryover occurs when analyte adheres to the probe and/or cuvette and contaminates a subsequent sample or cuvette in the testing sequence of the analyzer. Inadequate washing of the sample aspiration/ dispensing device and cuvette can cause this type of carryover. The Atellica® CI Analyzer uses independent integration and random-access sampling to process chemistry (CH) and immunoassay (IM) tests. The immunoassays performed on the IM portion of the analyzer utilizes a single-use, disposable sample tip and reaction cuvette which reduces sample carryover from IM to CH tests, whereas a reusable probe is used for CH assays—which is washed between sampling. This study evaluated the potential for carryover of CH samples from one sample to the next due to washing protocols on the Atellica® CI Analyzer in four independent real world clinical laboratories.

Methods

Carryover verification was performed as per CLSI GP31-A:2009 and Vericheck protocols. The study was conducted at four sites (UAE, France, Croatia, U.S.) using 2 serum samples prepared in 12 sample containers-9 for low (L) and 3 for high (H) concentrations of analyte. For glucose (GluH_3), UAE, L and H were 63 and 356 mg/dL (3.5 and 19.8 mmol/L), Croatia, 77.4 and 237.5 mg/dL (4.3 and 12.2 mmol/L), and U.S., 101 and 374 mg/dL (5.6 and 20.8 mmol/L). For creatine kinase (CK_L), France, L and H were 40 and 1400 U/L. Samples were tested in a defined sequence: L1, L2, L3, H1, L4, H2, L5, H3, L6, L7, L8, L9 and results analyzed for % carryover. Carryover was defined as average of L4, L5, L6 minus the average of L1, L2, L3, L7, L8, L9 divided by the average of H1, H2, H3.

Results

No significant carryover was observed at any site. Carryover % for UAE, France, Croatia, and U.S was 0.500%, -0.085%, -0.095%, and -0.095%, respectively.

Conclusions

When using high positive samples at concentrations at or greater than the upper limit of the clinical range, negligible CH sample carryover was found from the Atellica CI Analyzers in four independent real world clinical laboratory settings.
B-037 Zeon original coating technology COP microplates for cell imaging
Authors: Ishigami T , Adachi S , Funayama S , Kotagiri K , Ichimura N
Published: October 2024

Background

Zeon has developed high quality microplate made of Cyclo Olefin Polymer (COP) and special coat reagent for cell culture for Aurora microplates. The new 96 well-microplates have best-in-class properties for analysis of cultured cells can reduce the background of fluorescence images absolutely and excellent bottom flatness for observation of target. This enables to obtain high-precision data for cell analysis. Also, we developed Zeon original coating (ZOC) reagent specialized for COP surface. It is suitable for various cells including neurons derived from human induced pluripotent stem cells (iPSC), primary cells, general cell lines, and so on. ZOC coated COP microplates (ZOC plates) are fully-precoated and ready-to-use.

Methods

We evaluated the following points regarding ZOC plates. The cells used were derived from human iPSC.

[1. Storage stability] ZOC plates were stored at room temperature for a long time. After that, cardiomyocyte cells were cultured for 3 days on the plates. The cell adhesion and beat rate were evaluated.

[2. Neurite extension] Dopaminergic neurons were cultured for 14 days, then the neurite extension was evaluated. For comparison, a conventional poly-D-lysine* (PDL) coating was used.

[3. Lot-to-lot variation] COP microplates coated with different lots of ZOC reagents were prepared. Glutamatergic neurons were cultured for 14 days on those plates, then the neurite extension and cell morphology were evaluated. *The typical coating reagent applied for neuronal culture.

Results

[1.] ZOC plates stored over 12 months at room temperature showed excellent cell adhesion, and the beat rate was equivalent to prepared just before use.

[2.] ZOC plates tended to extend neurites more than PDL coated plates.

[3.] No variation between lots was observed on ZOC plates regarding the neurite extension and cell morphology.

Conclusions

We suggest applying the combination of high accuracy for cell imaging using COP microplates and high performance for cell culture using ZOC can provide high precision data at bioassay research. This would be a significant value for development of drug screening processes at pharmaceutical company.
B-048 Proposal of a new monitoring algorythm for patients with a HIV positive result with the Elecsys HIV DUO test
Authors: Castro Menendez J A , Leal Leyva Y Y , Garcia Gil A
Published: October 2024

Background

Worldwide HIV infections remain a public health problem, contributing significantly to morbidity. In Mexico we have had a progressive adaptation to the HIV epidemic providing free therapy and testing (4th generation tests) for the whole population in anbulatory centers for HIV/AIDS prevention and medical care. During 2022 Mexico reported 12,374 new cases. In the last decade there has been an increasing interest in the identification of patients HIV positives subgroups, based on disease progression and serological features. To date, 6 generations of tests for HIV detection have been developed, strongly reducing the window period for a positive result. Elecsys HIV Duo is a 5th generation immunological test that provides in a separate manner an antibody and antigen results for HIV detection. These results show a better insight of the serologycal status reducing the window period post infection. By using these tests we want to provide an early detection of new cases that gives a better prognosis for the patient and propose a new monitoring for patients with a positive antigen result but that have not yet produced antibodies. Such algorythm is currently lacking in the Mexican public health services.

Methods

A retrospective statystical analysis of HIV results of samples procesed with the Elecsys HIV DUO test during the january-april 2023 period in the National Reference Center of Salud Digna correlated with western blot results.

Results

The study population was 6405 patients, 6340 (98.98%) had a negative result in the test and 65 (1.02%) a positive test. Based in antibody/antigen results we categorized the positives results in three grupos: Group 1 Ab positive/Ag negative (16), group 2 Ab positive/Ag positive (32) and group 3 Ab negative/Ag positive (7). When correlated with the WB results all the patients from group 1 and 2 had a positive result while all patients in group 3 had a negative result.

Conclusions

Actually Mexican public health services do not have access to 5th generation HIV tests like the Elecsys HIV DUO test, therefore, public health profesional are not qualified to interprete these results and do not have a stated algorythm for patientes belonging to group 3. Therefore and based in our results we suggest 5th generation test (or superior if available) and propose a new patient monitoring algorythm with a molecular confirmatory test for viral detection. This proposal is strongly supported by the fact the we have a population that have not generated antibodys during the first weeks of post infection and will test negative in a serologycal confirmatory test like WB. Screening for patients in early infection stages is important because if we provide early therapy which may lead to infection reversal or a less impact in C4+ T cells numbers. Studies like this provide information for the generation of public politics in benefit of the population.
B-060 Prevalence of CCD and profilins antibodies in a seasonal allergy study
Authors: De Haro T , De Haro Romero T , González Cejudo T
Published: October 2024

Background

Allergen sources are complex and heterogeneous mixtures of proteins, which may contain from innocuous IgE-binding structures such as CCDs (cross-reactive carbohydrate determinants), which are not associated with symptoms, to a panallergen such as profilin, which is associated with multiple sensitizations to pollen and pollen-food-latex syndromes. CCDs are present in many plant and animal allergens and cause strong cross-reactivity due to the high similarity of their structure. They complicate the interpretation of positive in vitro diagnostic results because they do not associate symptomatology and, thats why, the determination of specific IgE antibodies against CCDs can provide useful information when positive IgE results are not consistent with the clinical picture. Profilins are ubiquitous proteins present in all eukaryotic cells and identified as allergens in pollen, latex and plant foods. The highly conserved structure explains the cross-reactivity of IgE antibodies against plant profilins and their designation as panallergens. As a panallergen, profilin is associated with multiple pollen sensitization and pollen-food-latex syndromes that the allergist must be aware of for accurate diagnosis and successful treatment of allergic diseases. We intend to evaluate the prevalence of these two markers with a new diagnostic tool, since profilins and CCDs are the substances most likely to cause in vitro cross-reactivity.

Methods

In our study 8,242 adults who presented rhinoconjunctivitis and visited their primary care center were included. The samples were analyzed in serum samples which were routinely processed in our laboratory with the EUROLINE Mediterranean Inhalation 2 (IgE) assay from Euroimmun® from February to July 2023, according to the manufacturer’s protocols. Patients with specific IgE levels ≥ 0.35 kU/L were identified as sensitized to allergen.

Results

A total of 8,242 patients were enrolled in this study, 5,173 women and 3,069 men, ranging in age from 15 to 65 years. Of these, 279 patients (3.39%) had antibodies against CCD, 570 (6.92%) antibodies against profilins and 183 (2.22%) had both. A higher prevalence of these antibodies was observed in men (CCD, n = 168, 5.47%; Profilins, n = 308, 10.04%) than in women (CCD, n = 111, 2.15%; Profilins, n = 262, 5.06%). Both markers were more frequently found in the presence of specific IgE antibodies to grass and weed pollens.

Conclusions

The possibility of detecting the presence of IgE antibodies against CCD and panallergenic profilins is useful to detect cross-reactivity in vitro, which could explain the differences found between laboratory results and patients' symptoms, helping the allergist to make a diagnosis with greater certainty and accuracy.
B-063 Changes in nonalcoholic fatty liver disease and M2BPGi due to lifestyle intervention in primary healthcare
Authors: Nah E , Choi Y , Park J
Published: October 2024

Background

A healthy lifestyle is the most important method for managing nonalcoholic fatty liver disease (NAFLD). Mac-2-binding protein glycosylated isomer (M2BPGi) has been suggested as a biomarker for NAFLD. This study aimed to determine the efficacy of personalized lifestyle interventions on NAFLD remission.

Methods

This single-arm intervention study recruited participants with NAFLD who underwent health checkups at seven health-promotion centers in five South Korean cities. Fatty liver diagnosis was based on ultrasonography (US). The 109 individuals were recruited for personalized lifestyle interventions of hypocaloric diets and exercise. The participants attended the lifestyle intervention programs once per month for the first 3 months, and once every 3 months for the subsequent 6 months. And phone-based self-monitoring were also provided monthly during the 3-month follow-up period. The primary outcome was NAFLD remission at month 12 as measured on US and magnetic resonance elastography. The secondary outcomes were the changes in metabolic factors and M2BPGi.

Results

The 108 individuals (62 males and 46 females; age 51.1±12.4 years, mean±standard deviation) were finally analyzed after the 12month intervention. Body mass index, waist circumference (WC), blood pressure, blood lipids (total cholesterol, triglycerides, and HDL-C), and fasting blood sugar levels were improved relative to baseline (all P<0.05). Fatty liver at or above the moderate grade according to US was decreased at month 12 relative to baseline (67.6% vs 50.9%) (P=0.002). M2BPGi levels decreased during the 12-month study period (P<0.001). M2BPGi levels were moderately correlated with hepatic fat fraction by magnetic resonance imaging (r=0.33, P=0.05). WC (OR=0.82, 95% CI=0.67-1.00, P=0.05) and HDL-C (OR=1.17, 95% CI=1.03-1.32, P=0.014) were associated with remission of fatty liver in the multivariate analysis.

Conclusions

The personalized lifestyle intervention was effective in improving fatty liver and metabolic factors, but not hepatic stiffness, in NAFLD.
B-067 Human IgE Monoclonal Antibodies as Unique Tools for Allergy Diagnostics
Authors: Filep S , Reid Black K , Smith B , Bermingham M , Pomes A , Chapman M
Published: October 2024

Background

Current blood-based tests for allergy diagnosis rely on human sera or plasma as the source of IgE antibody for method validation and calibration. Obtaining human samples with sufficiently high levels of IgE presents challenges for researchers and diagnostic companies. Here, we present a unique panel of allergen-specific human IgE monoclonal antibodies (mAb) that target inhalant, food and alpha-gal allergens and offer novel approaches for investigation of IgE responses and standardization of allergy diagnostic reagents and methods.

Methods

Human IgE mAb (n=30) targeting inhalant allergens from dust mite, cat and dog; food allergens from peanut, egg, milk, walnut and cashew, and galactose-α-1,3-galactose, were generated from B cells from symptomatic allergic patients with rhinitis, asthma, atopic dermatitis or red meat allergy using hybridoma technology. Total and allergen-specific IgE were quantified by ImmunoCAP (whole allergen ImmunoCAP were also performed). Allergen binding profiles and mAb titers were determined by two-site immunoassay (ELISA). Antibody purity was assessed using SDS-PAGE and Western Blot.

Results

SDS-PAGE showed single heavy and light chain bands under reducing conditions. Antibody recognition of target allergens was confirmed by Western Blot. Allergen-specific IgE mAb concentrations for inhalant allergens ranged: 9,000-37,000 kU/L for dust mite (n=3), 2,100-53,000 kU/L for cat (n=4), 5,300-21,000 for dog (n=2). Food allergens ranged: 16,300-49,400 kU/L for peanut (n=14), 15,700-81,000 for egg (n=2), 41,600 for milk (n=1), 7,100 for walnut (n=1) and 37,000 for cashew (n=1). Alpha-gal IgE monoclonals (n=2) ranged in concentration from 15,700-81,000 kU/L. The ratios of specific IgE to total IgE measured by ImmunoCAP vary by allergen and by individual human IgE mAb. Ratios for the inhalant mAb ranged from 0.04-1.05, for the food mAb the ratios ranged from 0.33-0.99. The two alpha-gal mAb showed relatively lower specific IgE levels (average ratio was 0.14; range=0.08-0.20).

Conclusions

Monoclonal IgE antibodies derived from patients presenting with allergic symptoms may provide a reliable replacement for human sera to investigate allergic disease. IgE mAb to inhalant and food allergens, and to the alpha-gal carbohydrate epitope, provide unique diagnostic markers and will offer improved accuracy for assessing patient allergy by standardizing diagnostic reagents and methods. Crystal structures of several of the human IgE mAb epitopes have recently been reported (Pomés et al, J Allergy Clin Immunol, 2024). The current panel of human IgE mAb can be expanded to include additional allergen targets, which may lead to the development of improved allergy therapeutics.
B-068 Plasma calprotectin as an index of draining tunnel formation in Hidradenitis suppurativa
Authors: Havelka A , Kanni T , Giamarellos-Bourboulis E J
Published: October 2024

Background

Hidradenitis suppurativa (HS) is a chronic, inflammatory skin disease which affects areas rich in apocrine glands. The main disease manifestations are inflammatory nodules (IN) and draining tunnels (dT). The course and progression of the disease are difficult to predict. There is currently lack of any broadly accepted biomarker which may inform on the activity of hidradenitis suppurativa (HS). Draining tunnel (dT) formation is the hallmark of HS in the affected skin. The existence of a biomarker which may predict dT formation before dTs appear in skin may become a tool for early biological treatment. Calprotectin, also known as S100 A8/A9 protein has been shown to be involved in various inflammatory conditions, including HS. Aim of this study was to evaluate circulating calprotectin as an index of dTs in the affected skin.

Methods

The study group consisted of 40 patients with confirmed HS and varying disease activity, assessed by number of dTs and International Hidradenitis Suppurativa Severity Score System (IHS 4 score) . Plasma samples were collected and stored at -80°C until analysis. Calprotectin was measured with GCAL® turbidimetric assay (Gentian AS, Norway).

Results

Fifteen patients had mild HS and 25 patients had moderate/severe HS. Positive association was found between circulating calprotectin and the absolute dT count (Pearson’s r2 = +0.750; p<0.0001) and the IHS4 (International HS 4 score) (Pearson’s r2 = +0.671; p<0.0001). Circulating calprotectin was significantly increased in patients with more than 2 dTs. Plasma calprotectin greater than 0.5 mg/L was associated with odds ratio 8.00 (1.87 to 34.22; p = 0.005) for 2 or more dTs.

Conclusions

Calprotectin is presented, for the first time, as a biomarker which indicates the formation of dTs in the skin. Its use as surrogate of early start of biologicals and response to treatment remains to be explored.
B-069 Characterization of a panel of antibodies as diagnostics for NASH and NAFL
Authors: van der Hoek A , Jaiu K , Gerhold J M
Published: October 2024

Background

Nonalcoholic fatty liver disease (NAFLD) is on the rise in Western industrialized countries and associated comorbidities like e.g. cirrhosis are a major cause of liver transplantations in the USA. In NAFLD, excess fat is accumulated in the liver whereas this build-up is not caused by alcohol. NASH (nonalcoholic steatohepatitis) and NAFL (nonalcoholic fatty liver) are two forms of NAFLD. NAFL represents a stage of the disease in which the liver does not exhibit major signs of inflammation or damage, while NASH does. NAFL does not necessarily progress to cause liver damage. NASH on the other hand may cause fibrosis and lead to cirrhosis with a risk of progression into liver cancer. Since NAFLD shows a prevalence of over 24-25% in both the USA and Europe, diagnosis moves more and more into focus. The current gold standard for NASH diagnosis is liver biopsy, and NAFLD as such can also be diagnosed by ultrasound and MRI, among others. A quest for biomarkers is ongoing to develop blood tests that allow NAFLD diagnosis by immunoassays for example. We have developed high affinity and specificity antibodies against four highly discussed marker proteins for NAFLD: RBP4 (Retinol Binding Protein 4), FGF21 (Fibroblast Growth factor 21), CHI3L1 (Chitinase-3-like protein 1) and PAI-1 (Plasminogen activator inhibitor-1). All four markers are discussed to be elevated in NAFLD patient serum.

Methods

We characterized the developed anti-RBP4 and anti-FGF21 antibodies via an in-house developed Chemiluminescence Immunoassay (CLIA).

Results

We have determined the limit of quantification (LoQ), limit of detection (LoD) and linear range of measurable concentrations for our antibody pairs. We further conducted spike and recovery experiments showing the precision of the developed antibodies against RBP4 and FGF21.

Conclusions

Together, we present antibody pairs for RBP4 and FGF21 suitable as a part of a diagnostic panel that has high potential to be applied in the diagnosis of NAFLD.
B-073 Analytical Performance of a Novel, Fully Automated Multiplexed Microarray Immunoassay Prototype for the Simultaneous Detection of Fifteen Autoantibodies Associated with Connective Tissue Diseases
Authors: Gomez G , Bijlsma D , Lukasik E , Hausmann M , Fischer C , Ghillani-Dalbin P , Miyara M , Moreau E
Published: October 2024

Background

There is currently a limited offer of automated multi-analyte tests for autoantibody testing in autoimmune connective tissue diseases (CTD). We assessed the analytical performance of a novel, single-use, multiplexed microarray immunoassay prototype (MosaiQ AiPlex CTDplus), used with the fully automated MosaiQ® system, for the simultaneous detection of fifteen autoantibodies associated with CTD, compared with selected CE-marked devices.

Methods

Banked human serum samples, characterized as reactive to ≥1 autoantibody or non-reactive were included. Reactive samples were characterized with FIDISTM Connective Profile (Theradiag) for CENP B, Jo-1, Ribosomal P, Scl-70, Sm, Sm/RNP, SS-A 60, TRIM21 (SS-A 52), SS-B and U1RNP; EliA CCP (Phadia) for CCP2, QUANTA Lite® Chromatin (Werfen) for Chromatin, QUANTA Flash® DFS70* (Werfen) for DFS70, Anti-dsDNA-IgG-ELISA (DRG) for dsDNA, and Immunodot (Euroimmun) for RNA polymerase III (RNAP III). For CCP2, only reactive samples were included. For the remaining analytes, non-reactive samples were characterized using immunofluorescence (ANA-Ro IgG FLUORESCENT HEp-2000®; Immuno Concepts, Germany) and ANAscreen ELISA (ORGENTEC-Diagnostika-GmbH, Germany). Each individual non-reactive sample was tested with the investigational device once; reactive samples were tested in duplicates. Positive percentage agreement (PPA) and negative percentage agreement (NPA) for individual analytes were calculated. For CCP2 only PPA is presented as no negative samples were included.

Results

Compared with relevant devices, the investigational prototype showed PPA ranging from 75% for chromatin to 99% for SS-A 60 and TRIM21. NPA ranged from 95% for chromatin, RNAP III, Scl-70 and Sm to 100% for DFS70, SS-A 60 and TRIM21. Performance details are shown in the Table.

Conclusions

The investigational prototype demonstrated substantial agreement with the compared CE-marked devices. Further studies will allow for expanded performance assessment of the investigational device. This fully-automated multiplexed platform has the potential to contribute to optimizing CTD evaluation by simplifying complex testing pathways and analyzing large number of samples per day.
B-075 Evaluation of patients with monoclonal gammopathy of undetermined significance using Optilite Freelite assays
Authors: Lakos G , Iliev V , Allen S , Rollins A , Foureau D , Bhutani M , McLendon K , Nooka A , Neish A , Birtwistle J
Published: October 2024

Background

Monoclonal gammopathy of undetermined significance (MGUS) is a pre-malignant disorder with a progression risk to multiple myeloma of 1% per year. Patients with MGUS may produce monoclonal intact immunoglobulins (Ig) and/or free light chains (FLC). The International Myeloma Working Group (IMWG) recommendations for the use of FLC measurements in MGUS are to establish the diagnosis of light chain only MGUS (LC-MGUS) and further to risk-stratify newly diagnosed MGUS. Once identified, MGUS is longitudinally monitored at intervals dependent on the risk of transformation. The IMWG recommends serum protein electrophoresis (SPEP) for monitoring of known MGUS with intact Ig, however others have published benefits of additionally monitoring with FLC. In this study, we evaluated the diagnostic sensitivity and specificity of the Optilite® Freelite® Kappa Free Kit and the Optilite Freelite Lambda Free Kit (The Binding Site, part of Thermo Fisher Scientific) in patients with MGUS, and the monitoring performance of Freelite results measured in serial samples from patients with clinically stable and progressive MGUS.

Methods

Serum samples from 234 patients with clinically diagnosed MGUS and 140 disease controls, taken at single time points, and serial samples (≥3) from 49 patients with MGUS were tested with the Freelite assays. Intact M-protein presence and isotype was confirmed in MGUS samples with immunofixation electrophoresis (IFE). In patients clinically diagnosed with LC-MGUS, the presence of detectable M protein on IFE was not required. The disease control cohort (non-MGUS) had disorders commonly associated with elevated polyclonal immunoglobulins (e.g autoimmune, liver, and renal diseases), confirmed by total IgG and/or IgA and/or IgM results and the absence of M-protein by IFE. Patients represented a diverse population from both European and US sources.

Results

The overall diagnostic sensitivity of the Optilite Freelite assays was 59.4%, calculated as the frequency of abnormal free light chain ratio (rFLC) in intact immunoglobulin MGUS, and abnormal rFLC with concurrent elevated involved FLC level in LC-MGUS. The diagnostic sensitivity was: 58.0% in non-IgM MGUS, 41.7% in IgM MGUS, 66.7% in biclonal MGUS and 100% in LC-MGUS. In intact (IgG/IgA/IgM) kappa MGUS, the diagnostic sensitivity was 67.5%, versus 41.5% in intact (IgG/IgA/IgM) lambda MGUS. The diagnostic specificity was 86.4%. In the 49 patients with serial samples, concentrations of involved FLC were stable in 93.3% of patients with clinically stable MGUS, whereas in patients with clinically progressive MGUS an increasing trend was seen in 50% of cases.

Conclusions

These results demonstrate that the Optilite Freelite Kappa and Lambda assays produce clinically relevant results that are useful in the evaluation of MGUS.
B-083 Sensitivity of the Sebia Hydragel 6 electrophoresis/immunofixation assay for detection of beta-2 transferrin in dilute cerebrospinal fluid (CSF): assessment by image analysis
Authors: Stickle D , Flerova E D
Published: October 2024

Background

Beta-2 transferrin (asialo transferrin) greater than 2-sialo transferrin is characteristic of cerebrospinal fluid (CSF). Our neurosurgery department recently requested availability of an in-house assay to evaluate post-surgical drainage fluids for presence of CSF. The Sebia Hydragel 6 beta-2 transferrin electrophoresis/immunofixation profile can be used as a qualitative (positive/negative) assay for this purpose. As a starting point for evaluation of the assay, we examined the sensitivity of the assay for detection of the beta-2 transferrin band in dilute CSF. This was performed by quantitative image analysis.

Methods

Assays were conducted according to manufacturer's instructions. Our proband sample was pooled CSF. Successive dilutions were by factors of 2 (1, 1/2, 1/4, 1/8, 1/16, 1/32). Electrophoresis/ immunofixation was performed simultaneously for these samples across lanes in 6-lane gels. We performed quantitative assessment of the electrophoretic bands for beta-2 transferrin by image analysis. The gel image was scanned to an electronic file (600 dpi png) using a desktop scanner. Grayscale pixel data were obtained using Python and the Python Image Library (PIL). Total grayscale integrals for staining of beta-2 transferrin bands were computed and compared across dilutions of the proband sample.

Results

For CSF dilutions, the assay was invariably visually positive down to dilutions of 1/16. By image analysis, this cutoff had signal/noise ratio (S/N) of 3.5. Across dilutions, band integrals were dependent on dilutions according to a simple hyperbola (representative example: see Figure).

Conclusions

The Sebia Hydragel 6 electrophoresis/immunofixation assay for qualitative assessment of presence of beta-2 transferrin was found to be suitable for unambiguous detection of beta-2 transferrin in CSF down to 1/16 dilution of CSF. By image analysis, the dose vs. response curve (density of staining for beta-2 transferrin across dilutions of CSF) was non-linear.
B-085 Optimizing Thyroglobulin Antibody Interference Thresholds to Minimize Turn-around Time
Authors: Patel S , Peck Palmer O M , Shurin M R , Wheeler S E
Published: October 2024

Background

Serum thyroglobulin (Tg) is a key biomarker used to detect disease recurrence in patients with differentiated thyroid cancers who have undergone complete thyroidectomy. Thyroglobulin antibodies (TgAb) can interfere with accurate measurement of serum Tg levels, producing falsely low results in automated immunoassays. Thus before measurement of Tg, TgAb levels are measured to determine if in-house immunoassays can be used or if the patient’s sample will need to be sent out for radioimmunoassay (RIA) or mass spectrometry methodologies, which are less sensitive to TgAb interference. We recently transitioned to new instrumentation and noted a sharp increase in Tg samples with TgAb interference that needed to be sent out when we used the limit of detection (LOD) of the Roche TgAb assay. This increases turnaround times for patients and increases laboratory costs. We hypothesized that the LOD of the Roche TgAb assay is below the level of interference in Tg immunoassays. To test this we compared the performance of TgAb immunoassays (IA) and Tg measurements by IA and RIA with the intention of proposing a more appropriate threshold for TgAb interference for the Roche TgAb assay.

Methods

We collected 121 serum specimens, which were above the LOD of the Roche TgAb (R-TgAb) assay, which is 10 IU/mL. These specimens were sent as part of standard care to USC for measurement of TgAb by Kronus (K-TgAb) and Tg by RIA. On these split samples we also measured Tg by Beckman Access (B-Tg). We compared the detection of TgAb by USC to varying thresholds of Roche TgAb measurements (10 IU/mL [LOD], 20 IU/mL, and 115 IU/mL [reference interval for autoimmunity]), and assessed the agreement between Tg measurements by Beckman IA and USC RIA for different Roche TgAb thresholds. Analyses were performed in Prism Graphpad using linear regression.

Results

Qualitatively, the concordance for R-TgAb to RIA (reference) at 10 IU/mL was 31%, at 20 IU/mL was 90%, and at 115 IU/mL was 80%. Quantitative agreement between the B-Tg immunoassay and RIA for the TgAb thresholds of: TgAb >10 IU/mL gave an R2=0.97 for Tg comparison, >20 IU/mL gave an R2=0.99, and >115 IU/mL R2=0.99.

Conclusions

Prior studies have shown that TgAb immunoassays can produce significant qualitative and quantitative differences. In our study, we attempt to validate the use of the Roche immunoassay with a cutoff of 20 IU/mL to minimize interference between Tg and TgAb in order to accurately detect recurrence of thyroid cancer. We found that a TgAb cutoff of 20 IU/mL we maintained acceptable agreement between Tg methods indicating no significant interference up to 20 IU/mL. With the TgAb cutoff of 20 IU/mL, we would be able to reduce sendouts for this cohort from 121 to 42, reducing sendouts by 65% and allowing a faster turnaround time for a significant proportion of our patients.
B-092 Effect of the Increased Specificity in a Recombinant anti-tTG Assay on a Celiac Disease Diagnostic Algorithm
Authors: Ferguson A , Morris C , Garg U
Published: October 2024

Background

Celiac disease is an autoimmune disease where ingestion of gluten results in damage in the small intestine. Celiac disease is estimated to affect 1 in 100 people worldwide, but only 30% of individuals are correctly diagnosed. If celiac disease is neither diagnosed nor treated, this can lead to additional health problems such as type 1 diabetes, multiple sclerosis, short stature, coronary artery disease and small intestine cancers. The celiac disease diagnostic algorithm at our institution begins with determination of IgA levels in the patient. If the patient is not IgA deficient (IgA > 7 mg/dL), an anti-tissue transglutaminase (anti-tTG) IgA level is determined. In patients who have anti-tTG IgA ≥ 4U/mL, an endomysial antibodies (EMA) IgA is performed by indirect immunofluorescence (IFA). Inova Diagnostics updated the reagent from human-derived native antigen to a more specific assay containing a recombinant antigen. We investigated the impact of this change on the downstream testing in our algorithm.

Methods

The new assay was implemented in August of 2023, and the monthly test counts and percentage of abnormal results were counted for both anti-tTG IgA and EMA IFA before and after the new assay was implemented.

Results

As shown in Figure 1., switching to the new recombinant anti-tTG reagent resulted in a decrease in the number of anti-tTG IgA results that were classified as abnormal. Subsequently, the abnormal samples that advanced in the algorithm for EMA IFA testing had a higher percentage of abnormal results, while the number of tests performed was lower.

Conclusions

Use of a more specific anti-tTG assay results in reduction of false positive results, and tests that continue through the celiac disease algorithm are more likely to represent a true diagnosis of celiac disease.
B-109 Six Sigma Analysis for Enhancing Biochemistry Internal Quality Control in a Private Laboratory in India
Authors: Patel T , Patel K
Published: October 2024

Background

This abstract outlines a comprehensive study on the application of Six Sigma methodologies to optimize internal quality control processes in a private biochemistry laboratory in India. Data collection was performed using Bio-Rad Assayed Chemistry Control on the Roche cobas 6000 instrument, facilitated by Bio-Rad Unity software. The study adheres to Clinical Laboratory Improvement Amendments (CLIA) 2024 guidelines, specifically focusing on Total Error (TEa).

Methods

The analytical performances of Cobas c501 insturment as analyse by running biochemical tests (urea, creatinine, uric acid, total bilirubin, AST, Amylase, Lipase, CK, LD, ALT, ALP, LDH, HDL-C, Ca2+, cholesterol, TG, HDL LDL, total protein, albumin, Na+, K+, and Cl−) and glucose on Cobas 400 were evaluated. Biases and CV were obtained from internal quality control programs using biorad unity software. The performance of each instrument was assessed using the σ scale; the benchmark performance was <3 σ (improvement needed); “world-class” quality was six sigma.

Results

Analytes such as “Alk PO4,” “ALT,” “Amylase,” “Bilirubin,” “CK,” “GGT,” “LD,” “Phosphorus,” “HDL,” “LDL Cholesterol,” “Triglycerides,” and “Uric acid” exhibited impressive sigma scores exceeding 6. Meanwhile, analytes including “Calcium,” “Total Cholesterol,” “Potassium,” “Total Protein,” “Urea,” and “Glucose” achieved sigma scores between 4 and 6, demonstrating noteworthy performance within acceptable bounds.

Conclusions

This study contributes valuable insights to laboratory management, specifically in the context of biochemistry testing in private Indian laboratories. The adoption of Six Sigma principles, coupled with adherence to CLIA 2024 guidelines and the utilization of advanced instrumentation and software, ensures compliance with regulatory requirements and fosters a culture of continuous improvement. This positions the laboratory to deliver high-quality and reliable results in the dynamic landscape of healthcare diagnostics.
B-118 Comparison of Statistical Methods, Chauvenet Test, and Interquartile Deviation for Urinary Sediment Inter Microscopists Evaluation
Authors: Ramadan-Boscolo D R , Banzato J M , Utiyama A H , de Andrade S S , Tufik S
Published: October 2024

Background

Comparison among microscopists is a quality process aimed at achieving comparability of analyses among collaborators who perform microscopy or manual analysis examinations, as in the case of urinary sediment examination. The current laboratory process evaluates red blood cell and white blood cell counts among microscopists within the same unit, aiming to identify discrepant counts through the application of statistical tools, with Chauvenet Statistics and Interquartile Deviation (IQD) being the most common in the literature. The purpose was to systematically compare Chauvenet Statistics and IQD as evaluation criteria for the comparison among microscopists for urinary sediment examination in our laboratories.

Methods

A total of 45 laboratory units within a total of 684 participants were analyzed throughout 2023. The results reported by microscopists from the same unit were evaluated using Chauvenet Statistics and IQD, calculating the adequacy rate for each type of assessment.

Results

Adopting IQD as the evaluation criterion, adequacy rates of 94.20% for red blood cell counts and 93.73% for white blood cell counts were observed. Using Chauvenet Statistics, the adequacy rates were 96.20% for red blood cells and 95.65% for white blood cells. This difference stems from 233 results identified as inadequate by the IQD method but deemed adequate by Chauvenet Statistics. In these cases, the results did not exhibit a normal distribution, approaching an expected distribution for categorical (discrete) variables. Additionally, 35 results identified as inadequate by Chauvenet Statistics were evaluated as adequate by the IQD method. These occurrences were observed when the reported results approached a normal distribution.

Conclusions

Regarding Chauvenet Statistics, IQD is a more efficient statistical tool for use as an evaluation criterion in the comparison among microscopists who perform red and white blood cell counts for urinary sediment examination in the selected laboratory units. However, the present study illustrated that each of the statistical tools will be more efficient in identifying outliers depending on the dataset analyzed. Therefore, it is important to consider the laboratory's epidemiology and population to decide which statistical tool is most suitable for each scenario.
B-119 Comparative performance of GPT-4 and CNV-ETLAI in extracting copy number variations from medical journals: Bridging the gap between large language models and specialized NLP tools in genomic data interpretation
Authors: Choi J
Published: October 2024

Background

Copy Number Variations (CNVs) are critical genetic markers in diversity and disease, yet their accurate extraction from medical literature remains challenging due to the complexity of genetic data. While specialized NLP models like CNV-ETLAI have been developed for this task, the advent of Large Language Models (LLMs) such as GPT-4 presents a potential alternative with broader applicability. This study evaluates the efficacy of GPT-4 against CNV-ETLAI in extracting CNVs from medical journal articles, aiming to enhance genetic research and clinical decision-making.

Methods

We configured GPT-4 to process and interpret medical journal PDFs, designing custom prompts for CNV information extraction. The performance of GPT-4 was benchmarked against CNV-ETLAI using a dataset of 146 true positive CNVs extracted from 23 journal articles. Performance metrics focused on accuracy in extracting CNVs from both text and tables, recognizing the importance of structured data interpretation in genomic analysis.

Results

CNV-ETLAI demonstrated superior accuracy, achieving a 98% success rate in CNV extraction, compared to GPT-4’s 49%. Specifically, CNV-ETLAI outperformed GPT-4 in table extraction accuracy (99% vs. 41.2%) and context extraction accuracy (96% vs. 63.2%). Despite GPT-4's lower performance, its capacity for improvement and adaptability was noted, indicating potential future applicability in medical data extraction.

Conclusions

The study highlights CNV-ETLAI's current superiority in extracting CNVs from medical texts, particularly in interpreting structured data like tables. However, the adaptability and potential for growth in LLMs like GPT-4 suggest they could soon become valuable tools for medical data extraction, offering a more versatile and powerful solution across a broader range of applications. The promise of LLMs, despite their current limitations, underscores the need for continued research and development in AI technologies for genomic data interpretation.
B-125 Laboratory Profiles for Diagnostic Workup of Acute Abdominal Pain in the Emergency Department: A Comparative Study of University and Non-University Acute Care Hospitals in Germany and Switzerland
Authors: Bauer I , Grossmann K , Risch L , Renz H , Risch M
Published: October 2024

Background

Laboratory medicine plays a fundamental role in patient care, especially in emergency departments where rapid diagnosis is critical to the efficient management of high patient volumes. With the aim of simplifying and standardizing laboratory requests, laboratory profiles have widely been established. This study uses the laboratory profile for “acute abdominal pain” as an example to illustrate the extent to which the laboratory profiles used in hospitals differ from each other and whether they are in line with current scientific recommendations.

Methods

Hospital data and laboratory profiles for “acute abdominal pain” from 18 hospitals, 9 from Germany and 9 from Switzerland, were analyzed and compared. The hospitals were distinguished by bed size, inpatient and outpatient case numbers, country affiliation and university level. In addition, an 'evidence-based laboratory profile' for acute abdominal pain was created based on current guidelines for the five most important causes of acute abdominal pain and compared with hospital laboratory profiles.

Results

German hospitals include significantly more laboratory parameters into acute abdominal pain profiles than Swiss hospitals (p = 0,0052). No significant differences in the number of laboratory parameters were found between university and non-university hospitals (p = 0,0507). There is a significant positive correlation between the hospital bed size and the number of laboratory parameters in the profiles (r = 0,5521). The number of inpatient cases in a hospital are also positively correlated with the number of laboratory parameters in the profiles (r = 0,4751). The number of outpatient cases ist not associated with the number of laboratory in the profiles parameters (r = 0,0599). The evidence-based laboratory profile for acute abdominal pain includes 21 laboratory parameters, but only two of them are collected by all investigated hospitals (bilirubin and ALAT).There were no significant differences in the fulfillment of the evidence-based laboratory profile between Germany and Switzerland, or between university or non-university status.

Conclusions

The study shows that there are quantitative and qualitative differences between the laboratory profiles for investigation of acute abdominal pain within the participating hospitals. In Germany and Switzerland there is no consensus on which analyses to include into a profile investigating acute abdominal pain. For increasing efficiency and accuracy of laboratory testing in in the diagnostic workup of acute abdominal pain it seems advisable to obtain consensus on what to reasonably include into a profile on a national or international level.