bluuebunny/crossref_metadata_2025_split huggingface dataset:
A-170 Hypereosinophilic Syndrome and t(5;15): Case Report and Literature Review
Authors: de Andrade S S , de Queiroz V C , Vicari P , Sthel V M , Perez B B , Figueira S H , Marcelino G A , do Prado G A , Ramadan-Boscolo D R , Tufik S 
Published: October 2024

Abstract

Background
Hypereosinophilic syndrome (HES) is a myeloproliferative disorder characterized by persistent eosinophilia (absolute eosinophil count &gt;1,500/mm3), classified into primary/clonal, reactive and idiopathic. After ruling out secondary causes of eosinophilia (parasitic infections, allergic conditions, vasculitis and collagenosis, drugs, eosinophilic lung disease, allergic gastroenteritis and metabolic conditions such as adrenal insufficiency), the main hypotheses become oncohematological diseases. Main entities to consider are myeloid/lymphoid neoplasms associated with eosinophilia and rearrangement of PDGFRA, FDGFRB, FGFR1 or PCM1-JAK2 and chronic eosinophilic leukemia. Conventional and molecular cytogenetics are the basis for classification and diagnosis of various hematological malignancies. This report describes a rare case of HES with t(5;15) (q33;q22).

Methods
A 29-year-old male patient was admitted with leukocytosis with the following exams: hemoglobin 6.5g/dL, hematocrit 21.7%, platelets 71,000/mm3, leukocytes 56,190/mm3 (60% neutrophils with a left shift to pro-myelocyte; 6% monocytes and 24% eosinophils), DHL 277 U/L, B12 vitamin 932 pg/mL, folic acid 1.72ng/mL, ferritin 913ng/mL. Non-reactive results were detected for antinuclear antibody, rheumatoid Factor, HIV, hepatitis B and C, syphilis and chagas disease. Bone marrow was hypercellular for age with absolute granulocytic predominance (G:E ratio 64:1) and eosinophilia (38%) in all maturational stages without dysplasia. Bone marrow flow cytometry did not detect neither increases in blasts nor anomalous immunophenotype. Bone marrow biopsy demonstrated 99% cellularity with normal blasts. Cytogenetics with translocation between the long arms of chromosome 5 and 15 in 15 of 20 metaphases analyzed - 46,XY,t(5;15)(q33;q22). No other molecular tests were available for this patient.

Results
In the case described t(5;15)(q33;q22) was found, which may be related to the TP53BP1::PDGFRB gene rearrangement. This breakpoint generates a chimeric oncoprotein similar to other tyrosine kinase fusions responsive to Imatinib. 53BP1 is a cellular DNA damage response component that binds wild type (non-mutant) p53. Oncogenesis occurs since the coiled-coil domains of TP53BP1 can mediate the homodimerization of PDGFRB and the constitutive activation of its tyrosine kinase activity; on the other hand, the DNA damage response of TP53BP1 may be disrupted. Gene rearrangements involving PDGFRA and FDGFRB are associated with response to tyrosine kinase inhibitors. Elevated vitamin B12 is a common finding in myeloproliferations with eosinophilia due to the increased production of haptocorrins, which binds to vitamin B12 in serum and in other tissues.

Conclusions
To our knowledge, in the literature there is only one described case of Myeloproliferative Neoplasia with Eosinophilia related to t(5;15)(q33;q22).
A-203 A Disconnect Between Detectable Patient Error Rates and QC Understanding
Authors: Brooks Z 
Published: October 2024

Abstract

Background
ISO 14971 defines “risk evaluation as the process of comparing the estimated risk against given risk criteria to determine the acceptability of the risk. Effective QC processes would detect if a stable acceptable error rate became unacceptable.

Methods
A survey was circulated through LinkedIn and Labvine. Forty respondents who included their credentials were included in the final analysis. Questions included:
1. What is the best choice for acceptable error rate when an analytical process is stable and in control, 2. If an analytical process “fails,” how many errors are you willing to report before the lab should stop reporting, and 3. “If an analytical process has a 5% stable error, how high would the error rate after failure need to become for a 1-2s rule to detect failure with one QC sample in a single QC run?”
This interactive poster will allow viewers to cast their vote for Question 4 and then reveal an LJ chart depicting a stable error rate of 5% with a shift to show the number of 1-2s failures with their chosen acceptable post-failure error rate.

Results
During stable operation, only 8% wanted zero errors while 38% voted that they would allow up to 5% errors;
After failure, 66% wanted to report zero errors and only 10% would allow 5% failure.
69% believed they would see a 1-2s failure in one run if the error rate rose to less than 50%; 19% believed it required a 100% error rate.

Conclusions
There is a strange disconnect where most believed it was acceptable to report more errors during stable operation than if the method failed.
There is a widespread misunderstanding of the logic that connects management of acceptable patient risk with the capabilities of statistical QC.
A-215 Feasibility of a unique homogenous assay for lipoprotein(a) particle number without bias from apo(a) size isoforms
Authors: Contois J , Albert A L , Nguyen R L , Datta P 
Published: October 2024

Abstract

Background
Lipoprotein(a) [Lp(a)] is an LDL-like particle with apolipoprotein(a) [apo(a)] covalently attached to apolipoprotein B. Elevated Lp(a) is associated with increased risk of atherosclerotic heart disease. The structural homology of Lp(a) and plasminogen and the variable number of apo(a) kringle 4 type 2 subunits (k4t2) between individuals, up to 51 repeats and a variable MW of 300-800 kDa, presents an analytical challenge. `Selection of calibrators of specific isoform size does not fix the problem, despite manufacturer’s claims to the contrary.

Methods
We have developed goat antisera to human apo(a)-specific peptides and a prototype immunoturbidimetric assay for Lp(a). We selected and synthesized 10 peptide sequences (15-30 aa each) distinct from the sequences found in k4t2 regions and plasminogen. Peptides were prepared attached to KLH (GenScript) and combined to immunize goats following a standard protocol including monthly boosters (Nittobo). Next, we assessed various sample volumes, reagent formulations and antisera dilutions to establish parameters on the Randox Imola analyzer. Cross-reactivity with recombinant k4t2 (40 mg/dL) and plasminogen (200 mg/dL, Athens Research) was assessed by comparison to the LOD (using delipidated plasma blank and low concentration samples).

Results
Titers to human Lp(a)) were positive to 1:6400 dilution of antisera, showing exceptional reactivity. Working formulation/parameters were 10 µL sample, 100 µL PBS, pH 7.4/PEG6000/detergent and 95 µL R2 (antisera). Sample/R1 was incubated for 5-minutes before addition of R2; reaction was monitored at 340 nm/37°C. LOD and linearity were also assessed. No cross-reactivity with k4t2 or plasminogen up to the concentrations tested was observed. Linearity to 330 nmol/L and LOD of 18 nmol/L were found.

Conclusions
The use of this antisera/assay eliminates the bias associated with apo(a) size polymorphisms and allows for the accurate measurement of Lp(a) particle number. More work is needed to optimize assay parameters and formulation and comprehensively validate assay performance.
A-219 Method Intercomparison Study between Atellica Solution® and CI® Analyzers for Total Cholesterol and Triglycerides
Authors: Ruiz-Alvarez M , Lapeña-Garcia S , Garcia-Valdecasas S , Costales-Lucia T , Moreno-Romero A , Molina-Blas S , Barrionuevo-Gonzalez M 
Published: October 2024

Abstract

Background
Method intercomparison is an essential requirement that every Laboratory must perform when considering a method or instrument change for a specific analyte.

Objective
verify if the results of two analytical techniques (Total Cholesterol and Triglycerides) measured by Atellica Solution® and CI® analyzers (Siemens Healthineers) are interchangeable.

Methods
40 patient samples were selected for each assay, with results distributed across the entire measurement range and analyzed by both instruments. Estatistical program Method Validator was used. To determine the degree of agreement between both instruments, Bland-Altman difference analysis and Passing-Bablok regression were performed. Statistics are expressed with their 95% confidence intervals.

Results
Total Cholesterol: After analyzing Bland-Altman differences, a systematic error was found as the mean difference between the results obtained by Atellica Solution® and CI 1900® was 13.8 with a 95% CI that does not include the null value. Regarding Passing Bablok analysis, a constant systematic difference was observed as the confidence interval of the intercept does not include the zero value, and also a proportional difference based on the 95% CI of the slope that does not include the value 1.• Triglycerides: After analyzing the Bland-Altman differences, a systematic error was found as the mean difference between the results obtained by Atellica Solution® and CI 1900® was -15.6, with higher results in Atellica Solution®. Regarding the Passing Bablok analysis, constant systematic differences were observed as the confidence interval of the intercept does not include the zero value, and also proportional differences based on the slope, whose 95% CI does not include the value 1

Conclusions
It´s concluded that Atellica Solution® and CI® analyzers are not interchangeable for Total Cholesterol and Triglycerides assays due to the presence of a constant systematic error. To use them on both instruments, it would be advisable to modify the reference values for both methods.
A-232 Characterize and reduce positive interference of hemolysis on Fungitell testing for (1, 3)-β-glucan (BDG) in serum
Authors: Zhang Y , Finkelman M , D’ordine R , Kowald G , Roy J 
Published: October 2024

Abstract

Background
Fungitell is an FDA cleared in vitro diagnostic product with an indication for use as an adjunct test for the diagnosis of invasive fungal infection. This assay has been established using a kinetic photometric method. A key aspect of this (1→3)-β-glucan (BDG) activated cascade reaction is its characteristic kinetic optical density profile. Interference which affects the BDG kinetic profile makes data analysis and interpretation difficult. This interference is roughly classified as being associated with hemolysis, icterus, lipemia (collectively HIL), and other non-specific factors.

Methods
In this study, we found that the use of 5% PEG-8000 supplemented serum (1:1 ratio of the volume of serum to 10% PEG-8000 solution with 0.1 M Tris-Cl, pH 7.4), heated at 80°C for 3.5 min, followed by centrifugation at 15,000 x g for 10 minutes (PEG-X Method) can largely eliminate the hemolysis-associated interference.

Results
The treated serum Fungitell BDG reaction curves are more like the ideal BDG kinetic profile. The recovery of introduced β-glucan has been shown to be acceptable (80-120%) for hemolyzed specimens.

Conclusions
This simple and rapid procedure shows promise as a pre-analytical treatment to reduce serum interference in the Fungitell β-glucan test for a substantial fraction of samples that produce currently unreportable results.
A-248 Assessment of Nanopore Sequencing for HIV Genotyping: A Comparative Study with Sanger Sequencing
Authors: Andrade M , Belmok A , Silva L , Vieira A , Vieira C , Velasco L , Jácomo R , Nery L 
Published: October 2024

Abstract

Background
The HIV drug resistance test, conducted through genotyping of the Human Immunodeficiency Virus (HIV), is a critical tool for assessing drug resistance in virus-infected patients and is commonly performed using Sanger Sequencing methodology (first-generation sequencing). Nanopore sequencing (third-generation sequencing) is a new technology that started to be commercialized a few years ago and has recently undergone significant updates with the new Q20+ chemistry. Therefore, this study aims to evaluate the potential use of nanopore sequencing (MinION) for HIV genotyping.

Methods
The genomic region encoding HIV protease and transcriptase was amplified by nested PCR using primer pairs modified with unique barcodes in the second round of amplification. Three libraries were assembled, including samples from 22 HIV-positive patients, one negative control (negative control patient), and one no-template control (NTC) per library, totaling 66 HIV-positive samples. The libraries were constructed using the SQK-LSK114 kit and sequenced on a MinION platform using the new Q20+ chemistry in flowcells or flongle R10.4.1. The sequences were assembled and analyzed using HIVdb-NGS (Stanford). Validation tests, such as accuracy and precision, were conducted following WHO recommendations for HIV genotyping. The same samples were also sequenced using the Sanger method for comparison.

Results
62 HIV sequences were generated by the MinION and analyzed in the described pipeline. The average sequence coverage per sample was 25,866 x. 41 sequences were classified as type B (63.08%), 12 as type C (18.46%), 8 as type F (12.31%), three as CRF02_AG (4.62%), and 1 as a recombinant B+C (1.54%). All identified subtypes were consistent with those found by the Sanger method. The average similarity between the sequences of each sample (MinION vs Sanger) was 98.68% (minimum 94.96%, maximum 99.95%). Basecalling similarity at relevant positions (associated with resistance) was above 98% in 59 out of 62 samples (95.16%), meeting the acceptance criteria recommended by the WHO.

Conclusions
Our results suggest that nanopore sequencing (utilizing Q20+ chemistry) can replace Sanger sequencing for HIV genotyping, which is used to monitor the effectiveness of anti-HIV drugs. This transition will allow for a reduction in test costs in laboratories with high demand for HIV drug resistance testing. Additionally, the use of modified primers proposed here eliminates the need for commercial barcodes, resulting in reduced processing time and improved throughput.
A-281 Comparison of C-reactive Protein, Procalcitonin, and Presepsin in Blood and Synovial Fluid as Diagnostic Biomarkers for Septic Arthritis
Authors: Choi H , Choi J , Jeon S , Kim M , Seon J , Kee S 
Published: October 2024

Abstract

Background
Early detection and prompt treatment are crucial for managing septic arthritis (SA). However, diagnosing SA at an early stage can be challenging since the symptoms are non-specific, and there is no definitive diagnostic test available. This study aimed to investigate the potential of biomarkers as a reliable diagnostic tool for SA.

Methods
A prospective study was conducted to collect residual blood and synovial fluid samples from patients who underwent blood sampling and arthrocentesis for differential diagnosis due to suspected SA. The collected samples were analyzed for C-reactive protein (CRP), procalcitonin, and presepsin. The data obtained from the analysis were subjected to statistical analysis.

Results
From January to November 2023, 58 patients were enrolled. They comprised 24 men and 34 women with a median age of 71.1 (interquartile range, IQR 64.0-79.3) years. Twenty-one patients were diagnosed with SA and 37 with non-SA, including osteoarthritis (n=2), osteonecrosis (n=1), meniscus tear (n=1), abscess (n=1), contusion (n=2), and mechanical complications of implants (n=30). There were no significant differences in age or sex distribution between the two groups. The median blood and synovial fluid CRP levels were 4.24 and 1.85 mg/dL, respectively, in the SA group and 0.24 and 0.11 in the non-SA group. The median blood and synovial fluid procalcitonin levels were 0.0383 and 0.0403 ng/mL in the SA group and 0.0358 and 0.0488 in the non-SA group. The median blood and synovial fluid presepsin levels were 307 and 1389 pg/mL in the SA group and 192 and 959 in the non-SA group. Except for blood procalcitonin, all showed statistically significant differences between the SA and non-SA groups. In the ROC curve analysis, blood CRP displayed the highest area under the curve (AUC) of 0.795 with a 0.47 mg/dL cut-off value, 90.5% sensitivity, and 64.9% specificity. Synovial fluid CRP displayed similar results of 0.786 AUC, 0.30 mg/dL cut-off value, 85.0% sensitivity, and 71.4% specificity. Blood and synovial fluid procalcitonin had low AUCs of 0.532 and 0.659, respectively, and were insufficient to differentiate between SA and NS groups. The AUC (0.696 vs. 0.704), sensitivity (76.2% vs. 76.2%), and specificity (67.6% vs. 62.2%) of blood and synovial fluid presepsin were similar; however, synovial fluid showed a significantly higher cut-off value, 4.5 times that of blood (237 pg/mL vs. 1087 pg/mL).

Conclusions
Procalcitonin was not a reliable marker for distinguishing between SA and non-SA, regardless of whether it was measured in blood or synovial fluids. However, CRP and presepsin proved useful markers when measured in either blood or synovial fluid. In particular, when measuring presepsin in synovial fluid, an optimal cut-off value is needed.
A-290 Prevalence and Antimicrobial Resistance Pattern of Microorganisms Recovered From Bloodstream Infections From Public Hospitals of São Paulo City, Brazil
Authors: de Andrade S S , Nurmberger J M , Inoue F M , Lobo A P , Ramadan-Boscolo D R , Tufik S 
Published: October 2024

Abstract

Background
Nosocomial Bloodstream Infections (nBSI) are an important cause of morbidity and mortality. Brazilian studies concerning bloodstream infections are important tools that can bring specific issues related to antimicrobial resistance. The objective of this study was to describe the frequency and antimicrobial susceptibility profile of bacterial isolates from BSI in patients admitted to public hospitals in 2022 and 2023.

Methods
This study was a retrospective analysis from 12 hospitals (January 2022 to December 2023) based on BSI isolates laboratory data detected by the clinical microbiology department of a medical diagnostic laboratory. The identification and susceptibility tests were performed using MALDI-TOF mass spectrometry (Vitek MS) and Vitek®2 systems, according to the manufacture’s recommendations. Coagulase Negative Staphylococci (CoNS) BSI was only considered in the presence of at least two sets of positive blood cultures with the same organism.

Results
During the period (January 2022 to December 2023) a total of 19,588 microorganisms isolated from BSI were obtained from 12 hospitals. The frequency of organism group was Gram-positive (10,707; 54.7%), Gram-negative (7,874; 40.2%), yeasts (961; 4.9%) and filamentous (46; 0.2%). The order of prevalence (top ten) organisms was CoNS (5,149; 26.3%); Staphylococcus aureus (2,769; 14.1%); Klebsiella spp. (2,355; 12.0%); Escherichia coli (1,301; 6.6%); Acinetobacter spp. (1,293; 6.6%); Streptococcus spp. (1,125; 5.7%); Enterococcus spp. (1,112; 5.7%); Candida spp. (937; 4.8%); Pseudomonas spp. (749; 3.8%) and Proteus spp. (553; 2.8%). Extended spectrum beta-lactamase rates for E. coli and Klebsiella pneumoniae were 31.6% (268/848) and 74.9% (1027/1371), respectively. The resistance to carbapenems was detected mainly in 74.1% (661/892) of Acinetobacter spp. isolates, 60.7% (832/1371) of Klebsiella pneumoniae; 24.5% (114/465) of P. aeruginosa; 9.7% (34/349) of Proteus spp. and 1.4% (12/828) of Escherichia coli. Antimicrobial resistance was observed mainly in 87.2% of CoNS (2831/3247) and 44.5% of S. aureus (743/1669) to methicillin; 22.9% (146/637) of Enterococccus spp. to vancomycin.

Conclusions
The frequency of Klebsiella spp. isolates from nBSI has progressively increased and effective infection control measures should be taken to prevent them from spreading. The rates of antimicrobials resistance reported for Klebsiella pneumoniae, Acinetobacter spp., CoNS, S. aureus and Enterococcus spp. reinforce the high resistance rates to antimicrobials in São Paulo city.
A-326 Testing for Multidrug-Resistant Acinetobacter baumannii with a Microfluidic Device
Authors: Gärtner C , Treffon J , Kreuchauff J , Heppner B , Isserstedt-John N , Klemm R R , Omengo B , Mellmann A 
Published: October 2024

Abstract

Background
Multidrug-resistant (MDR) Acinetobacter baumannii in hospitals contribute to high mortality rates and entail expensive infection control procedures. Conventionally, species and resistance determination of MDR bacteria is performed by mass spectrometry and labor-intensive phenotypical assays in clinical microbiology laboratories. Some institutions also apply time-consuming and expensive whole genome sequencing for in-depth resistance and transmission analysis. In contrast, faster and more cost-efficient MDR typing can be achieved by on-site application of real-time PCR-based point-of-care tests (POCTs) that target species- and resistance-specific genes and enable rapid screening of large sample sizes.

Methods
To construct a real-time PCR-based POCT system for species and resistance determination of MDR A. baumannii, we selected two species-specific genes (rpoB, OXA-51-like) and 18 resistance genes found in MDR A. baumannii (OXA-23-like, OXA-24/40-like, OXA-51-like, OXA-58-like, OXA-143-like, mcr, ADC, NDM, GIM, VIM, IMP, GES, PER, TEM, SHV, VEB, CTX-M, KPC) as targets. For each gene, one to six sets of primers and fluorophore-labeled probes that amplify conserved genetic regions were designed and their functionality was tested in vitro. Parallel, a fully automated system comprising microfluidic chips and a bench-top analyzer able to perform DNA extraction and real-time PCR starting from swab eluate was developed.

Results
All oligonucleotides tested so far reliably detected 100 to 100,000 copies of DNA extracted from MDR bacteria in single- and multiplex experiments in vitro. Tests with the microfluidic system showed that lysis of different bacterial species could be accomplished via sonication on a membrane located on the microfluidic chip. In addition, chip implementation of all PCR reagents like lyophilized master mix as well as primers and probes was successful and can be stored on chip at least one year. So far, the system delivered detectable fluorescence signals within one hour when performing test runs with bacteria eluted from swabs.

Conclusions
A real-time PCR-based Point-of-Care screening platform was developed for the detection of antibiotic-resistant A. baumannii from various patient sample materials starting directly from swabs, which detects colonization/infection and at the same time the relevant resistance genes within about an hour.