scholarly journals Automated Determination of Serum α1-Antitrypsin by Antitryptic Activity Measurement

2009 ◽  
Vol 55 (3) ◽  
pp. 513-518 ◽  
Author(s):  
Denis Roche ◽  
Alexandra Mesner ◽  
Malik Al Nakib ◽  
Frederic Leonard ◽  
Philippe Beaune
Keyword(s):  
Large Scale ◽  
Reference Interval ◽  
Reference Population ◽  
Activity Measurement ◽  
Trinitrobenzene Sulfonic Acid ◽  
Serum Samples ◽  
Automated Assay ◽  
Automated Method ◽  
A1at Deficiency ◽  
Antitryptic Activity

AbstractBackground: α1-Antitrypsin (A1AT) deficiency is currently detectable by protein immunoassay, phenotyping, and genotyping of the S and Z mutations, but no fully automated method for standard biochemical analyzers is yet available. Here, we present a method that measures the antitryptic activity in serum. This method is rapid, automated, and allows the easy evaluation of a large cohort of patients.Methods: Our automated assay involves determining serum antitryptic capacity on the Olympus AU 400 autoanalyzer by using trypsin and succinylated gelatin as substrate in the presence of trinitrobenzene sulfonic acid. The results are expressed as a percentage of inhibition of the reaction of trypsin with succinylated gelatin. After we performed analytical validation studies and reference-interval determination based on serum samples from 120 healthy persons, we tested the assay on deidentified samples from 120 patients with various pathologies (primarily pulmonary) of unexplained origin and normal A1AT concentrations and phenotypes.Results: The analysis rate was up to 120 samples per hour. Intraassay CVs ranged from 3.1%–16.2%, and interassay CV was 7.5%. The reference population showed mean (SD) 58.4 (6.7)% inhibition. The detection limit was 9.5% inhibition. The 120 studied patients displayed significantly lower mean activity than 120 healthy individuals (P < 0.0001).Conclusion: This assay is stable, reliable, and easily automated by use of open-system analyzers, allowing for the rapid evaluation of patients. After further validation on a larger randomized cohort, this new approach should function as a useful method to explore A1AT deficiency, especially in large-scale studies.

scholarly journals Active B12: A Rapid, Automated Assay for Holotranscobalamin on the Abbott AxSYM Analyzer

2008 ◽  
Vol 54 (3) ◽  
pp. 567-573 ◽  
Author(s):  
Jeff Brady ◽  
Lesley Wilson ◽  
Lynda McGregor ◽  
Edward Valente ◽  
Lars Orning
Keyword(s):  
Vitamin B12 ◽  
Serum Vitamin ◽  
Vitamin B12 Deficiency ◽  
Reference Interval ◽  
Cross Reactivity ◽  
Total Serum ◽  
Serum Samples ◽  
Automated Assay ◽  
Microparticle Enzyme Immunoassay ◽  
B12 Deficiency

Abstract Background: Conventional tests for vitamin B12 deficiency measure total serum vitamin B12, whereas only that portion of vitamin B12 carried by transcobalamin (holotranscobalamin) is metabolically active. Measurement of holotranscobalamin (holoTC) may be more diagnostically accurate for detecting B12 deficiency that requires therapy. We developed an automated assay for holoTC that can be used on the Abbott AxSYM immunoassay analyzer. Methods: AxSYM Active B12 is a 2-step sandwich microparticle enzyme immunoassay. In step 1, a holoTC-specific antibody immobilized onto latex microparticles captures holoTC in samples of serum or plasma. In step 2, the captured holoTC is detected with a conjugate of alkaline phosphatase and antiTC antibody. Results: Neither apoTC nor haptocorrin exhibited detectable cross-reactivity. The detection limit was ≤0.1 pmol/L. Within-run and total imprecision (CV ranges) were 3.4%–5.1% and 6.3%–8.5%, respectively. Assay CVs were <20% from at least 3 pmol/L to 107 pmol/L. With diluted serum samples, measured concentrations were 104%–114% of the expected values in the working range of the assay. No interference from bilirubin, hemoglobin, triglycerides, erythrocytes, rheumatoid factor, or total protein was detected at expected (abnormal) concentrations. A comparison of the AxSYM Active B12 assay with a commercial RIA for holoTC yielded the regression equation: AxSYM = 0.98RIA + 4.7 pmol/L (Sy x, 11.4 pmol/L; n = 204). Assay throughput was 45 tests/h. A 95% reference interval of 19–134 pmol/L holoTC was established with samples from 292 healthy individuals. Conclusions: The AxSYM Active B12 assay allows rapid, precise, sensitive, specific, and automated measurement of human holoTC in serum and plasma.

Automation and validation of a MALDI-TOF MS (Mass-Fix) replacement of immunofixation electrophoresis in the clinical lab

2021 ◽  
Vol 59 (1) ◽  
pp. 155-163
Author(s):  
Mindy Kohlhagen ◽  
Surendra Dasari ◽  
Maria Willrich ◽  
MeLea Hetrick ◽  
Brian Netzel ◽  
...  
Keyword(s):  
Limit Of Detection ◽  
Turnaround Time ◽  
Automated System ◽  
Serum Samples ◽  
Maldi Tof Ms ◽  
Maldi Tof ◽  
Automated Method ◽  
Positive Specimen ◽  
Specimen Identification ◽  
Tof Ms

AbstractObjectivesA matrix assisted laser desorption ionization time of flight mass spectrometry (MALDI-TOF MS) method (Mass-Fix) as a replacement for gel-based immunofixation (IFE) has been recently described. To utilize Mass-Fix clinically, a validated automated method was required. Our aim was to automate the pre-analytical processing, improve positive specimen identification and ergonomics, reduce paper data storage and increase resource utilization without increasing turnaround time.MethodsSerum samples were batched and loaded onto a liquid handler along with reagents and a barcoded sample plate. The pre-analytical steps included: (1) Plating immunopurification beads. (2) Adding 10 μl of serum. (3) Bead washing. (4) Eluting the immunoglobulins (Igs), and reducing to separate the heavy and light Ig chains. The resulting plate was transferred to a second low-volume liquid handler for MALDI plate spotting. MALDI-TOF mass spectra were collected. Integrated in-house developed software was utilized for sample tracking, driving data acquisition, data analysis, history tracking, and result reporting. A total of 1,029 residual serum samples were run using the automated system and results were compared to prior electrophoretic results.ResultsThe automated Mass-Fix method was capable of meeting the validation requirements of concordance with IFE, limit of detection (LOD), sample stability and reproducibility with a low repeat rate. Automation and integrated software allowed a single user to process 320 samples in an 8 h shift. Software display facilitated identification of monoclonal proteins. Additionally, the process maintains positive specimen identification, reduces manual pipetting, allows for paper free tracking, and does not significantly impact turnaround time (TAT).ConclusionsMass-Fix is ready for implementation in a high-throughput clinical laboratory.

scholarly journals European Biological Variation Study (EuBIVAS): within- and between-subject biological variation estimates for serum thyroid biomarkers based on weekly samplings from 91 healthy participants

2021 ◽  
Vol 0 (0) ◽  
Author(s):  
Michela Bottani ◽  
Aasne K. Aarsand ◽  
Giuseppe Banfi ◽  
Massimo Locatelli ◽  
Abdurrahman Coşkun ◽  
...  
Keyword(s):  
Large Scale ◽  
Thyroid Stimulating Hormone ◽  
Biological Variation ◽  
Healthy Individuals ◽  
Serum Samples ◽  
Free Triiodothyronine ◽  
Performance Specifications ◽  
Study Population ◽  
Variance Homogeneity ◽  
Healthy Participants

Abstract Objectives Thyroid biomarkers are fundamental for the diagnosis of thyroid disorders and for the monitoring and treatment of patients with these diseases. The knowledge of biological variation (BV) is important to define analytical performance specifications (APS) and reference change values (RCV). The aim of this study was to deliver BV estimates for thyroid stimulating hormone (TSH), free thyroxine (FT4), free triiodothyronine (FT3), thyroglobulin (TG), and calcitonin (CT). Methods Analyses were performed on serum samples obtained from the European Biological Variation Study population (91 healthy individuals from six European laboratories; 21–69 years) on the Roche Cobas e801 at the San Raffaele Hospital (Milan, Italy). All samples from each individual were evaluated in duplicate within a single run. The BV estimates with 95% CIs were obtained by CV-ANOVA, after analysis of variance homogeneity and outliers. Results The within-subject (CV I ) BV estimates were for TSH 17.7%, FT3 5.0%, FT4 4.8%, TG 10.3, and CT 13.0%, all significantly lower than those reported in the literature. No significant differences were observed for BV estimates between men and women. Conclusions The availability of updated, in the case of CT not previously published, BV estimates for thyroid markers based on the large scale EuBIVAS study allows for refined APS and associated RCV applicable in the diagnosis and management of thyroid and related diseases.

scholarly journals A non-targeted LC–MS metabolic profiling of pregnancy: longitudinal evidence from healthy and pre-eclamptic pregnancies

Metabolomics ◽  
2021 ◽  
Vol 17 (2) ◽  
Author(s):  
Tiina Jääskeläinen ◽  
◽  
Olli Kärkkäinen ◽  
Jenna Jokkala ◽  
Anton Klåvus ◽  
...  
Keyword(s):  
Large Scale ◽  
Scale Effects ◽  
Late Pregnancy ◽  
Metabolite Profile ◽  
Adverse Outcomes ◽  
Liquid Chromatography Mass Spectrometry ◽  
Targeted Metabolomics ◽  
Serum Samples ◽  
Healthy Women ◽  
Maternal Metabolism

Abstract Introduction Maternal metabolism changes substantially during pregnancy. However, few studies have used metabolomics technologies to characterize changes across gestation. Objectives and methods We applied liquid chromatography–mass spectrometry (LC–MS) based non-targeted metabolomics to determine whether the metabolic profile of serum differs throughout the pregnancy between pre-eclamptic and healthy women in the FINNPEC (Finnish Genetics of Preeclampsia Consortium) Study. Serum samples were available from early and late pregnancy. Results Progression of pregnancy had large-scale effects to the serum metabolite profile. Altogether 50 identified metabolites increased and 49 metabolites decreased when samples of early pregnancy were compared to samples of late pregnancy. The metabolic signatures of pregnancy were largely shared in pre-eclamptic and healthy women, only urea, monoacylglyceride 18:1 and glycerophosphocholine were identified to be increased in the pre-eclamptic women when compared to healthy controls. Conclusions Our study highlights the need of large-scale longitudinal metabolomic studies in non-complicated pregnancies before more detailed understanding of metabolism in adverse outcomes could be provided. Our findings are one of the first steps for a broader metabolic understanding of the physiological changes caused by pregnancy per se.

scholarly journals Low seroprevalence rates of Zika virus in Kuala Lumpur, Malaysia

2019 ◽  
Vol 113 (11) ◽  
pp. 678-684 ◽  
Author(s):  
I-Ching Sam ◽  
Magelda Montoya ◽  
Chong Long Chua ◽  
Yoke Fun Chan ◽  
Andrew Pastor ◽  
...  
Keyword(s):  
Zika Virus ◽  
Large Scale ◽  
Binding Assay ◽  
Seroprevalence Rate ◽  
Protective Effects ◽  
Kuala Lumpur ◽  
Serum Samples ◽  
Denv Serotypes ◽  
Population Immunity ◽  
Low Incidence

Abstract Background Zika virus (ZIKV) is believed to be endemic in Southeast Asia. However, there have been few Zika cases reported to date in Malaysia, which could be due to high pre-existing levels of population immunity. Methods To determine Zika virus (ZIKV) seroprevalence in Kuala Lumpur, Malaysia, 1085 serum samples from 2012, 2014–2015 and 2017 were screened for anti-ZIKV antibodies using a ZIKV NS1 blockade-of-binding assay. Reactive samples were confirmed using neutralization assays against ZIKV and the four dengue virus (DENV) serotypes. A sample was possible ZIKV seropositive with a ZIKV 50% neutralization (NT50) titre ≥20. A sample was probable ZIKV seropositive if, in addition, all DENV NT50 titres were <20 or the ZIKV NT50 titre was >4-fold greater than the highest DENV NT50 titre. Results We found low rates of possible ZIKV seropositivity (3.3% [95% confidence interval {CI} 2.4 to 4.6]) and probable ZIKV seropositivity (0.6% [95% CI 0.3 to 1.4]). Possible ZIKV seropositivity was independently associated with increasing age (odds ratio [OR] 1.04 [95% CI 1.02 to 1.06], p<0.0001) and male gender (OR 3.5 [95% CI 1.5 to 8.6], p=0.005). Conclusions The low ZIKV seroprevalence rate, a proxy for population immunity, does not explain the low incidence of Zika in dengue-hyperendemic Kuala Lumpur. Other factors, such as the possible protective effects of pre-existing flavivirus antibodies or reduced transmission by local mosquito vectors, should be explored. Kuala Lumpur is at high risk of a large-scale Zika epidemic.

Pediatric reference interval verification for endocrine and fertility hormone assays on the Abbott Alinity system

2021 ◽  
Vol 0 (0) ◽  
Author(s):  
Mary Kathryn Bohn ◽  
Siobhan Wilson ◽  
Alexandra Hall ◽  
Khosrow Adeli
Keyword(s):  
Clinical Decision Making ◽  
De Novo ◽  
Reference Interval ◽  
Clinical Decision ◽  
Reference Intervals ◽  
Healthy Children ◽  
Confidence Limits ◽  
Test Results ◽  
Serum Samples ◽  
Abbott Architect

Abstract Objectives The Canadian Laboratory Initiative on Pediatric Reference Intervals (CALIPER) has developed an extensive database of reference intervals (RIs) for several biomarkers on various analytical systems. In this study, pediatric RIs were verified for key immunoassays on the Abbott Alinity system based on the analysis of healthy children samples and comparison to comprehensive RIs previously established for Abbott ARCHITECT assays. Methods Analytical performance of Alinity immunoassays was first assessed. Subsequently, 100 serum samples from healthy children recruited with informed consent were analyzed for 16 Alinity immunoassays. The percentage of test results falling within published CALIPER ARCHITECT reference and confidence limits was determined. If ≥ 90% of test results fell within the confidence limits, they were considered verified based on CLSI guidelines. If <90% of test results fell within the confidence limits, additional samples were analyzed and new Alinity RIs were established. Results Of the 16 immunoassays assessed, 13 met the criteria for verification with test results from ≥ 90% of healthy serum samples falling within the published ARCHITECT confidence limits. New CALIPER RIs were established for free thyroxine and prolactin on the Alinity system. Estradiol required special considerations in early life. Conclusions Our data demonstrate excellent concordance between ARCHITECT and Alinity immunoassays, as well as the robustness of previously established CALIPER RIs for most immunoassays, eliminating the need for de novo RI studies for most parameters. Availability of pediatric RIs for immunoassays on the Alinity system will assist clinical laboratories using this new platform and contribute to improved clinical decision-making.

A fully automated method of human identification based on dental panoramic radiographs using a convolutional neural network

2021 ◽  
Author(s):  
Young Hyun Kim ◽  
Eun-Gyu Ha ◽  
Kug Jin Jeon ◽  
Chena Lee ◽  
Sang-Sun Han
Keyword(s):  
Neural Network ◽  
Convolutional Neural Network ◽  
High Speed ◽  
Large Scale ◽  
Oral Surgery ◽  
Human Identification ◽  
Running Time ◽  
Automated Method ◽  
Image Characteristics ◽  
Proposed Model

Objectives: This study aimed to develop a fully automated human identification method based on a convolutional neural network (CNN) with a large-scale dental panoramic radiograph (DPR) dataset. Methods: In total, 2,760 DPRs from 746 subjects who had 2 to 17 DPRs with various changes in image characteristics due to various dental treatments (tooth extraction, oral surgery, prosthetics, orthodontics, or tooth development) were collected. The test dataset included the latest DPR of each subject (746 images) and the other DPRs (2,014 images) were used for model training. A modified VGG16 model with two fully connected layers was applied for human identification. The proposed model was evaluated with rank-1, –3, and −5 accuracies, running time, and gradient-weighted class activation mapping (Grad-CAM)–applied images. Results: This model had rank-1,–3, and −5 accuracies of 82.84%, 89.14%, and 92.23%, respectively. All rank-1 accuracy values of the proposed model were above 80% regardless of changes in image characteristics. The average running time to train the proposed model was 60.9 sec per epoch, and the prediction time for 746 test DPRs was short (3.2 sec/image). The Grad-CAM technique verified that the model automatically identified humans by focusing on identifiable dental information. Conclusion: The proposed model showed good performance in fully automatic human identification despite differing image characteristics of DPRs acquired from the same patients. Our model is expected to assist in the fast and accurate identification by experts by comparing large amounts of images and proposing identification candidates at high speed.

Pediatric reference intervals for 1,25-dihydroxyvitamin D using the DiaSorin LIAISON XL assay in the healthy CALIPER cohort

2018 ◽  
Vol 56 (6) ◽  
pp. 964-972 ◽  
Author(s):  
Victoria Higgins ◽  
Dorothy Truong ◽  
Nicole M.A. White-Al Habeeb ◽  
Angela W.S. Fung ◽  
Barry Hoffman ◽  
...  
Keyword(s):  
Children And Adolescents ◽  
Critical Role ◽  
Reference Interval ◽  
Reference Intervals ◽  
Biologically Active ◽  
Specific Reference ◽  
Healthy Children ◽  
Serum Samples ◽  
Dihydroxyvitamin D ◽  
1,25 Dihydroxyvitamin D

Abstract Background: 1,25-dihydroxyvitamin D (1,25(OH)2D), the biologically active vitamin D metabolite, plays a critical role in calcium and phosphate homeostasis. 1,25(OH)2D is measured to assess calcium and phosphate metabolism, particularly during periods of profound growth and development. Despite its importance, no reliable pediatric reference interval exists, with those available developed using adult populations or out-dated methodologies. Using the fully automated chemiluminescence immunoassay by DiaSorin, we established 1,25(OH)2D pediatric reference intervals using healthy children and adolescents from the CALIPER cohort. Methods: Serum samples from healthy subjects (0 to <19 years) were analyzed for 1,25(OH)2D using the DiaSorin LIAISON XL assay and age-specific reference intervals were established. The Mann-Whitney U-test was used to determine seasonal differences. Pooled neonatal and infantile samples were quantified using liquid chromatography tandem mass spectrometry (LC-MS/MS) to determine if elevated concentrations during the first year of life may be attributed to cross-reacting moieties. Results: Three reference interval age partitions were required with highest levels in subjects 0 to <1 year (77–471 pmol/L), which declined and narrowed after 1 year (113–363 pmol/L) and plateaued at 3 years (108–246 pmol/L). 1,25(OH)2D concentration was not significantly affected by seasonal variation or sex. Elevated 1,25(OH)2D concentrations in neonatal and infantile samples may be the result of an interfering substance. The absence of 3-epi-1,25-dihydroxyvitamin D in the pooled samples makes it unlikely to be the interfering moiety. Conclusions: Pediatric reference intervals for 1,25(OH)2D were established to improve test result interpretation in children and adolescents. 1,25(OH)2D is elevated in a proportion of neonates and infants, which may be the result of a cross-reacting moiety.

scholarly journals Mining Connected Vehicle Data for Beneficial Patterns in Dubai Taxi Operations

2018 ◽  
Vol 2018 ◽  
pp. 1-8 ◽  
Author(s):  
Raj Bridgelall ◽  
Pan Lu ◽  
Denver D. Tolliver ◽  
Tai Xu
Keyword(s):  
Large Scale ◽  
Statistical Tests ◽  
Poor Quality ◽  
Connected Vehicle ◽  
Cleaning Method ◽  
Mobility Data ◽  
Automated Method ◽  
Vehicle Data ◽  
Chi Squared ◽  
Trip Production

On-demand shared mobility services such as Uber and microtransit are steadily penetrating the worldwide market for traditional dispatched taxi services. Hence, taxi companies are seeking ways to compete. This study mined large-scale mobility data from connected taxis to discover beneficial patterns that may inform strategies to improve dispatch taxi business. It is not practical to manually clean and filter large-scale mobility data that contains GPS information. Therefore, this research contributes and demonstrates an automated method of data cleaning and filtering that is suitable for such types of datasets. The cleaning method defines three filter variables and applies a layered statistical filtering technique to eliminate outlier records that do not contribute to distributions that match expected theoretical distributions of the variables. Chi-squared statistical tests evaluate the quality of the cleaned data by comparing the distribution of the three variables with their expected distributions. The overall cleaning method removed approximately 5% of the data, which consisted of errors that were obvious and others that were poor quality outliers. Subsequently, mining the cleaned data revealed that trip production in Dubai peaks for the case when only the same two drivers operate the same taxi. This finding would not have been possible without access to proprietary data that contains unique identifiers for both drivers and taxis. Datasets that identify individual drivers are not publicly available.

A Preliminary Assessment of Vitamin K1 Intakes and Serum Undercarboxylated Osteocalcin Levels in 11–13 Year Old Irish Girls

2006 ◽  
Vol 76 (6) ◽  
pp. 385-390 ◽  
Author(s):  
Collins ◽  
Cashman ◽  
Kiely
Keyword(s):  
Vitamin K ◽  
Large Scale ◽  
Bone Growth ◽  
Food Composition ◽  
Undercarboxylated Osteocalcin ◽  
Mineral Density ◽  
Serum Samples ◽  
Old Irish ◽  
Dietary History ◽  
Food Composition Data

Low vitamin K1 intakes have been associated with low bone mineral density in women and reduced bone turnover in girls. No European data exist on the relationship between vitamin K1 and serum undercarboxylated osteocalcin (ucOC), an indicator of K1 status in adolescents. The aim of the current study was to assess intakes of vitamin K1 in relation to serum ucOC status in Irish girls. A detailed dietary history method, which measured habitual intakes from a typical 14-day period, was used to estimate vitamin K1 intakes in 18 girls aged 11–13 years. Recently compiled and validated food composition data for vitamin K1 were used to determine vitamin K1 intakes. An enzyme immunoassay was used to measure ucOC in fasting serum samples. The mean (± SD) intake of vitamin K1 in the girls was 72.4 μg/day (SD 34.4). Vegetables (particularly broccoli, composite dishes, and lettuce) contributed 53% of total vitamin K1 intakes. Thirty-Seven percent of the girls failed to meet the current U.S. adequate intake for adolescents of 60 μg/day vitamin K1. Serum ucOC levels were inversely related to body weight-adjusted vitamin K1 intakes, controlling for energy intake (partial correlation r = –0.538; p = 0.026). The data indicate that large-scale studies to examine relationships between vitamin K1 (and green vegetable) intakes and bone growth and development in adolescents are warranted.

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