bluuebunny/crossref_metadata_2025_split huggingface dataset:
B-035 Maintaining the Analytical Performance of Calcium and Phosphorus in 24-hour Urine Without Addition of 6n hcl in the Pre-Analytical Process
Authors: Canali D G , Zampieri G , Motta C M , Oliveira D C , Rosin C , Argolo S V , Satsuma W 
Published: September 2023

Abstract

Background
The measurements of Calcium (Ca) and Phosphorus (P) in 24-hour urine samples are associated with the evaluation of hyperparathyroidism, renal tubular acidosis, and monitoring of Falconi Syndrome and Nephrolithiasis. The aim is to demonstrate the maintenance of analytical performance without the addition of 6N hydrochloric acid (HCl) in the pre-analytical process.

Methods
The study was divided into stages and carried out for both analytes. In the first stage, correlation tests were performed on 40 samples with and without HCl within the range of linearity. Imprecision was processed at 3 clinically relevant concentrations for 5 days, repeated 5 times. In the second stage, stability testing of Ca and P was performed on 7 samples over 5 days, without HCl (pH 5–6.7), with HCl (pH &lt; 3), alkalized (pH &gt; 8), and neutral (pH 7–7.5). In the last stage, interferent testing was performed on 286 samples, where pH was measured after collection and after acidification with 200uL of HCl. A pH meter and pH strips were used to measure pH, and a Siemens® - Advia 2400 equipment was used for analytical measurement of Ca and P.

Results
In the correlation results, the variance test for both analytes resulted in F-test: 1.01 &lt; 1.69, the difference analysis was less than 2.02, correlation coefficient and Bias% of 1.00 and 0.55 for Ca and 0.99 and 0.27 for P, respectively. Total error (TE) obtained for Ca was 9.69 and 9.41, lower than the allowed TE for both analytes. The repeatability and intralaboratory imprecision of the three levels of each analyte were less than 13.8% for Ca and 9.0% for P. There was 100% correlation of the results when the analyses were performed with samples at pH up to 7; at pH values &gt; 7, the concentrations decreased by 60%. All 268 samples with different pH values, subjected to 200uL of HCl, were suitable for Ca and P measurement with pH below 5.7.

Conclusions
Samples with a pH greater &gt;or = to 7 necessarily need to be acidified with HCl 6N. There is no analytical alteration if all samples in the routine of urinary Ca and P are submitted to 200uL of HCl 6N, even the most alkaline samples. The non-provision of preservative containers to patients reduces the risk of accidents, costs, and the need for legal licenses for handling in healthcare facilities.
B-047 Performance Evaluation of Enzymatic HbA1c Assay on Newly Launched Mindray BS-600M Chemistry Analyzer
Authors: Li M , Xie W , Zeng Y , Xu A , Ji L , Dai J 
Published: September 2023

Abstract

Background
HbA1c has important reference value in risk stratification, diagnosis, therapeutic monitoring and drug selection for diabetic patients. Therefore, the ability to detect HbA1c has attracted much attention. This study was designed to evaluate the performance of the enzymatic HbA1c assay on a newly launched Mindray BS-600M chemistry analyzer, which can be used to measure whole blood samples directly after mixing without centrifugation.

Methods
Total 309 clinical samples (EDTA anticoagulation) were analyzed using Mindray BS-600M (enzymatic method), Tosoh G8 (HPLC method) and Roche c501 (immunoassay method) analyzers for the performance comparison. And also, the analytical performance of BS-600M, such as precision, accuracy, linearity, interference of conventional interferents, hemoglobin variants, hemoglobin derivatives and common drugs, were evaluated using other fresh whole blood samples, eight frozen calibrators (IFCC) and forty frozen venous whole blood samples (NGSP).

Results
The within-run and between-run precision of the enzymatic method for HbA1c determination were less than 1% and linearity of HbA1c (%, NGSP unit) was excellent from 2.96% to 20.23%. The deviations between the results of 48 accuracy samples (IFCC and NGSP) and target values were within ±5%, and the sigma metrics of HbA1c at the high and low concentration levels were between 9.9∼19.0, which belongs to the "world-class" level. The results were excellently correlated with those obtained by Tosoh HLC -723 G8 and Roche Cobas c501 (r = 0.997, 0.992). The regression equations of HbA1c was y = 0.979x + 0.005 (n = 198) between BS-600M and G8 and y = 0.934x + 0.317 (n = 112) between BS-600M and c501. The deviation of interference from conventional interferents (bilirubin, Intralipid, vitamin C), hemoglobin variants (HbS, HbC, HbD, HbE), hemoglobin derivatives (LAlc, CarHb, AcetylatedHb), and commonly used antipyretic analgesics and hypoglycemic drugs were all within ±6%, except for the high level HbF (&gt;10%); and also, BS-600M cannot be applied to severe anemia patients (HGB &lt; 45 g/L), which may lead to falsely high HbA1c results.

Conclusion
The Mindray BS-600M proved to be a robust and reliable method for HbA1c measurement, and was suitable for routine clinical chemistry or hematology laboratory use. In addition, attention or comment need to be added for samples having high HbF concentrations and severe anemia patients in reporting HbA1c% result.
B-056 Performance Characteristics of a New, Fully Automated Planar Microarray Immunoassay for the Detection of IgG Auto-Antibodies Against Centromere Protein B (CENP-B)
Authors: Daubrosse C , Ghillani-Dalbin P , Mercier V , Pasion Galvan R , Bijlsma D , Lukasik E , Botti V , Gomez G , Moreau E , Hausmann M , Ginocchio C C , Miyara M 
Published: September 2023

Abstract

Background
Detection of auto-antibodies against Extractable Nuclear Antigens (ENA), including CENP-B, is key in identification of autoimmune (AI) diseases. Many current test methods are manual, slow, labor intensive or do not facilitate the simultaneous detection of several auto-antibodies, that could reduce time to provide comprehensive results to clinicians. We report the performance characteristics of MosaiQTM CENP-B (Quotient Suisse, Eysins, CH), a new planar microarray immunoassay designed for use with the fully automated, continuous random access, high throughput MosaiQ system for qualitative serological detection of CENP-B IgG auto-antibodies. CENP-B assay is one parameter of a multiplexed MosaiQ connective tissue disease panel in development.

Methods
MosaiQ CENP-B microarrays were prepared by printing CENP-B antigens onto epoxy-silane glass chips and assembled into magazines for processing on the MosaiQ 125 instrument (Fig. 1). Primary patient serum tubes and reagents are loaded on the instrument and automatically processed to generate immunoassay spot signals, interpreted by the instrument using a proprietary image analysis algorithm. Anonymized human serum samples (Hôpital Pitié Salpetrière, Paris, France), characterized as CENP-B auto-antibodies non-reactive or reactive with ≥1 CE-marked devices, were included in the study.

Results
After discordant analysis, MosaiQ system correctly identified 96/99 samples characterized as reactive by comparators and all 199 non-reactive samples: positive percent agreement 97.0% (95% CI, 91.4%, 99.4%), negative percent agreement 100% (95% CI, 98.2%, 100%), overall percent agreement 99% (95% CI, 97.1%, 99.8%). A reproducibility and repeatability evaluation showed, respectively, overall agreement of 99.7% and 100%.

Conclusion
MosaiQ CENP-B microarray provides a high level of clinical concordance with other CE-marked assays for detecting CENP-B auto-antibodies. The MosaiQ System has the potential to advance AI disease testing by increasing laboratory efficiency and productivity, with the ability to multiplex the detection of various auto-antibodies on a single microarray, and a capacity to automatically process thousands of samples per day.
B-059 Allergen-specific IgE Measurement for Peanut Ara h 2*
Authors: Chuang T , Ready N , Rowlands M , Chamberlain J , Hammer D , Sommer M , Hovanec-burns D 
Published: September 2023

Abstract

Background
Legumes such as peanuts are a major source of food allergies among children and adults. Ingestion is the primary route of exposure, although inhalation may also trigger an allergic response in patients with known hypersensitivity. Allergic reactions to peanut can include atopic dermatitis, angioedema, vomiting, diarrhea, and anaphylaxis. Several allergenic peanut proteins have been identified and linked with variable severity of patient symptoms. Ara h 2 is one of the proteins uniquely associated with severe peanut allergy. In this study, we evaluate the analytical performance of the Ara h 2 allergen containing reagent for use in the IMMULITE® 2000 3gAllergy® Specific IgE assay.

Methods
Serum samples were collected from 13 patient samples with documented histories of tree nut or legume-related allergies. The samples were analyzed for specific IgE reactivity to a commercially available recombinant Ara h 2 (rAra h 2) using the IMMULITE 2000 3gAllergy Specific IgE assay (Siemens Healthcare Diagnostics Inc., Tarrytown, NY). Precision, linearity, limit of detection, and endogenous interference were assessed.

Results
Repeatability and within-lab precision at all levels tested were less than 10% over a 20-day study. Regression analysis was performed following linearity assessment and resulted in a slope 95% CI of 0.9251–1.0492 (range 0.3–24 kU/L). Limit of blank and limit of detection were 0.02 kU/L and 0.08 kU/L, respectively. Endogenous interference testing showed &lt;10% difference when spiked with hemoglobin (1000 mg/dL), intralipid (3000 mg/dL), and unconjugated bilirubin (40 mg/dL).

Conclusion
rAra h 2 allergen containing reagent for use in the IMMULITE 2000 3gAllergy Specific IgE assay is sensitive and capable of quantitatively measuring IgE in patients with clinical history of nut allergy.
*Assay under development. Not available for sale. Future availability cannot be guaranteed.
B-097 Storage Stability of HbA1c in Whole Blood Tubes at Room Temperature on the Abbott Alinity c Clinical Chemistry System
Authors: Berman M , Wang X , Cheng P , Kulasingam V 
Published: September 2023

Abstract

Background
Hospital laboratories frequently receive requests to order additional tests on an existing specimen. For this reason, laboratories routinely save specimens after initial testing has been completed for a defined period of time. The current literature lacks specific information regarding HbA1c stability in whole blood and the effects of red blood cell settling. In addition, literature references are not inclusive of various conditions which can cause gaps in data collection such as samples in queue for prolonged periods. To obtain information on prolonged storage of whole blood tubes, HbA1c was assessed for storage at room temperature (RT) (20–25°C) for up to 4 hours on the Alinity c system. In addition, potential carryover from a whole blood to a serum sample was assessed.

Methods
A minimum of 40 whole blood sample pools (approx. 8 mLs) were prepared from remnant samples. Sample pools were mixed, aliquoted, and stored at RT on the Alinity c system. The samples were tested as a single replicate continuously for up to 4 hours without mixing (minimum of 20 cycles) and compared to the baseline sample values. Carryover from a whole blood to a glucose serum sample was assessed after 4 hours continuous testing of whole blood to mimic a high volume testing laboratory. The stability of each HbA1c sample and carryover into a glucose sample were evaluated by comparing the mean absolute or percent difference from the respective sample baseline measurement. The allowed deviation from baseline was 5%.

Results
After 4 hours at RT, the whole blood cells had settled to approximately 50% from the top of the tube. The red blood cell settling had no significant impact on the results. Within sample precision for the 20 cycles of repetitive testing was &lt;/ = 1%. The 48 individual samples showed insignificant difference from the baseline value, with a mean change of 1.0% (Range 0–3.2%). Sample carryover from the whole blood samples to a serum glucose sample was not significant (2.71 Baseline vs 2.64 first replicate).

Conclusion
This study demonstrated that the storage stability of HbA1c is stable for up to 4 hours at RT on the Alinity c system.
B-137 Development of a Novel Software System for Providing Real-Time Laboratory Test Information to Clinicians at Point of Order Entry
Authors: Douglas C S , Zaydman M 
Published: September 2023

Abstract

Background
Clinicians depend on an increasingly complex set of laboratory tests to aid in the diagnosis and monitoring of patients. Staying informed about the proper usage of these tests requires active effort, especially as test offerings continually evolve. To aid in test decision making, institutions often implement some form of decision support, including: grouping of common “order sets”, “best practice alert” popups, and web-based “lab catalogs”. For many institutions, lab catalogs provide the most comprehensive information on available tests, but typically exist separate from the interface used to place test orders, and thus suffer from underutilization in actual clinical practice.

Methods
We developed a novel, open-source software to closely integrate our institution’s lab catalog at the point of order entry within our electronic medical record (EMR) system. The software provides an immediate pop-up window anytime a clinician needs additional information while placing orders. The software also provides an independent, customizable search function that helps steer clinicians away from commonly mis-ordered tests. Both functions are immediately available at the point of order entry, without requiring the user to leave the EMR or otherwise interrupt their normal workflow. We conducted a small usability pilot with a focus group of clinicians, using the System Usability Scale (SUS) (1).

Results
Our software reduced the number of clicks and keystrokes needed to access detailed lab information from 27 to 2. The focus group (n = 6) rated the product favorably on the SUS, with average rating 89 ± 9 out of 100.

Conclusion
This study demonstrates proof-of-concept for a novel software decision support tool that provides instant access to lab information at the point of order entry. Future work will focus on evaluating utility in a real-world clinical pilot.
B-186 Investigating LC Column Selectivity for 19 Steroids in a Clinical Research Setting
Authors: Freige M , Marin S J 
Published: September 2023

Abstract

Background
Steroids are a class of structurally related endogenous compounds. Several have the same chemical formula (and hence the same m/z) and must be chromatographically separated for accurate identification and quantitation by LC-MS/MS. Here, six bonded silica solid core or “core-shell” LC columns (see Table 1) were evaluated to determine selectivity and optimum separation conditions for a 19 analyte steroid panel for clinical research. The optimum column for the separation of estrone and estradiol was also established.

Methods
Analytical reference standards (Cerilliant®, Round Rock, TX) were used to prepare diluted samples for injection. Six different core-shell 2.6 µm, 50 × 2.1 mm LC columns (Kinetex™, Phenomenex, Torrance, CA) were evaluated (see Table 1). Detection employed a 7500 triple quadrupole LC-MS/MS equipped with an ESI source used in positive and negative mode (SCIEX®, Framingham, MA) coupled to an Agilent® 1290 liquid chromatograph (Agilent Technologies, Santa Clara, CA).

Results
The traditional C18 column worked best for the complete steroid clinical research panel. Estrogens had the best separation on the biphenyl phase, which demonstrated the highest level of selectivity for estrone and estradiol. The phenyl-hexyl column did not sufficiently separate all steroid isomers in the clinical research panel, however it was sufficient to separate estrone and estradiol.

Conclusion
The proposed HPLC conditions and the core-shell 2.6 μm 50 × 2.1 mm C18 column provided the best separation for 19 steroids with a fast 8 minute run time. For research focused on estrogens, the core-shell biphenyl column yielded the best separation.
B-196 LC/MS-MS Quantification of Myo-inositol: A Novel Biomarker for Kidney Disease
Authors: Omosule C , Blair C , Herries E , Farnsworth C , Dietzen D , Gaut J 
Published: September 2023

Abstract
Currently accepted clinical standards for assessing kidney function include the measurement of creatinine for calculating estimated glomerular filtration rates (eGFR) and albumin to creatinine ratios. However, creatinine, a product of muscle creatine metabolism, is an imperfect proxy for renal function because it is subject to non-renal interferences. There is an unmet need for extremely sensitive and precise biomarkers for evaluating renal function in patients given that acute kidney injury (AKI) and chronic kidney disease (CKD) are linked to high mortality, morbidity, and contribute considerably to health care expenditures.Myo-inositol oxygenase (MIOX) is a novel biomarker limitedly expressed in the proximal renal tubules. MIOX concentrations are elevated in mice with renal injury and in critically ill patients with acute kidney injury (AKI), and increases are observed 54 h prior to changes in serum creatinine are observed. Importantly, MIOX facilitates the conversion of myo-inositol to glucuronic acid. We developed a robust LC-MS/MS method to quantify myo-inositol in human plasma and serum, hypothesizing that following renal injury (acute and chronic), plasma concentrations of myo-inositol will increase, and will correlate with increases in serum creatinine concentrations.In patients with stable renal function, the central 95% distribution of plasma myo-inositol concentrations was 16.61–44.2 µM (Median 27.7 µM, 95% CI: 26.7–28–7). No sex-based differences in plasma myo-inositol were observed. We also observed excellent correlation between plasma myo-inositol and serum creatinine concentrations (r = 0.84, 95% CI: 80–88), as well as excellent sensitivity (99%) and specificity (100%) at a myo-inositol concentration of 50.22 µM for differentiating patients with stable kidney function from those with CKD. Patients with CKD exhibited higher plasma myo-inositol concentrations (P value &lt; 0.0001) with a central 95% percentile of 51.2–309.1 µM (Median 104.1 µM, 95% CI: 92.3–113.6).These results imply that myo-inositol concentrations increase in accord with renal tubular injury and MIOX buildup in patients with kidney dysfunction, making it a potential biomarker for renal impairment.
B-212 Application of Annexin A5Coated Beads for Collection of Extracellular Vesicles and Their Content RNA as Liquid Biopsy for Lung Cancer
Authors: Mei-Chia W , Guan-Yu G , Chih-Liang W , Pi-Yueh C , Chiuan-Chian C 
Published: September 2023

Abstract

Background
Extracellular vesicles (EVs) carry molecules derived from cells, such as RNA. Measurement of tumor-derived extracellular RNA (exRNA) in EVs from plasma can be a promising approach for the development of blood-based cancer detection. However, the lack of an easy method for EVs separation limits its application. The aim of this study is to establish a procedure for EV collection and exRNA detection for non-small-cell lung cancer (NSCLC) diagnosis and monitoring.

Methods
Annexin A5-coated magnetic beads (ANX beads) were developed for EVs isolation. Next, we established a procedure for EV separation and exRNA detection from fluidic specimens using the ANX beads. Then blood samples from 22 NSCLC patients and 20 healthy controls were collected to evaluate the procedure and we detected different RNAs in the samples.

Results
We evaluated the ability of ANX beads in EVs separation and found that the beads could capture large and small EVs and could increase RNA concentration. Then we detected different RNAs in the samples and found that CK19 mRNA, MALAT1 lncRNA, and miR155 microRNA had good performance to differentiate NSCLC from healthy controls.

Conclusion
In conclusion, this assay protocol could separate EVs to enrich RNA concentration. The EVs separation employed was captured by magnetic beads which could be adopted into an automatic nucleic acid extract system to conveniently concentrate and extract nucleic acid from EVs.
B-228 Partition-free Digital Polymerase Chain Reaction: A Novel Approach to Enable Rapid Turnaround Time With Digital Accuracy
Authors: Erb P , Myrick J , Hensley M , Schreiber J , Sundberg S 
Published: September 2023

Abstract

Background
Partition-free digital PCR (dPCR) is a new approach developed to provide absolute quantification in less than 10 min. Partition-free eliminates the need for droplets or physical wells that need to be filled and sealed. The technique relies on rapid thermal cycling and microfluidics to limit diffusion and enable the identification of individual amplicon clusters originating from a single target nucleotide template. Rapid heating and cooling were achieved with a thermal cycling engine comprised of a broadband LED with a sapphire block that is both a light guide and heatsink. Sample is distributed through a custom microfluidic cartridge containing 64 microchannels for optical quantification.

Methods
The feasibility of partition-free dPCR was tested with primers targeting MTHFR c.665. A titration was performed on three 4X serial dilutions of cell line genomic DNA in triplicate. Fluorescent profiles were generated for all 64 microchannels from an image captured after 35 PCR cycles, where amplicon clusters were identified. Fluorescent peaks of the amplicon clusters were counted and quantified via a custom analysis algorithm, which uses Poisson statistics to calculate concentration.

Results
The quantification results obtained from the partition-free dPCR system were compared to concentration measurements taken via a Bio-Rad QX200 platform. Concentrations of 69.4 ± 0.8, 15.4 ± 1.5, and 3.3 ± 0.6 copies/µL (R2 = 0.9997) were measured on the partition-free system, and concentrations of 67.8 ± 3.6, 17.2 ± 1.1, and 2.8 ± 0.2 copies/µL (R2 = 0.9995) were measured on the Bio-Rad QX200.

Conclusion
It has been demonstrated that absolute quantification can be achieved in less than 10 min with a partition-free dPCR platform, and shows results comparable to a QX200 droplet dPCR system. The partition-free rapid dPCR technology described herein allows for the exploration of new avenues in molecular diagnostics, with the potential to bring absolute quantification of dPCR to a Point-of-Care setting.
B-259 Exocas-2 Rapid and Pure Isolation of Exosomes by Anionic Exchange using Magnetic Beads
Authors: Bae M , Shin S 
Published: September 2023

Abstract

Background
Extracellular vesicles (EVs) are considered essential biomarkers in liquid biopsies. Despite intensive efforts aimed at employing EVs in a clinical setting, workable approaches are currently limited owing to the fact that EV-isolation technologies are still in a nascent stage. This study introduces a magnetic bead-based ion exchange platform for isolating EVs called ExoCAS-2 (exosome clustering and scattering). Owing to their negative charge, exosomes can easily adhere to magnetic beads coated with a polycationic polymer. Owing to the features of magnetic beads, exosomes can be easily processed via washing and elution steps and isolated with high purity and yield within 40 min.

Methods
Cationic salt is coated on the surface of magnetic beads with carboxyl group surface residues. Inject the cation coated magnetic bead (ExoCAS) into the plasma solution and incubate in 4°C for 30 minutes. By charge interaction, negative charged EVs are attached to the cationic beads. And then, pH 6 washing buffer and 1M NaCl elution buffer were used for EV isolation.

Results
The present results confirmed the isolation of exosomes through analyses of size distribution, morphology, surface and internal protein markers, and exosomal RNA. Compared with the commercially available methods, the proposed method showed superior performance in terms of key aspects, including operation time, purity, and recovery rate.

Conclusion
This highlights the potential of this magnetic bead-based ion exchange platform for isolating exosomes present in blood plasma.