| 1. |
- Aljadi, Zenib, et al.
(författare)
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Microfluidic Immunoaffinity Basophil Activation Test for Point-of-Care Allergy Diagnosis
- 2019
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Ingår i: Journal of Applied Laboratory Medicine (JALM). - : American Association for Clinical Chemistry. - 2475-7241 .- 2576-9456. ; 4:2, s. 152-163
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Tidskriftsartikel (refereegranskat)abstract
- Background: The flow cytometry-based basophil activation test (BAT) is used for the diagnosis of allergic response. However, flow cytometry is time-consuming, requiring skilled personnel and cumbersome processing, which has limited its use in the clinic. Here, we introduce a novel microfluidic-based immunoaffinity BAT (miBAT) method. Methods: The microfluidic device, coated with anti-CD203c, was designed to capture basophils directly from whole blood. The captured basophils are activated by anti-FceRI antibody followed by optical detection of CD63 expression (degranulation marker). The device was first characterized using a basophil cell line followed by whole blood experiments. Weevaluated the device with ex vivo stimulation of basophils in whole blood from healthy controls and patients with allergies and compared it with flow cytometry. Results: The microfluidic device was capable of capturing basophils directly from whole blood followed by in vitro activation and quantification of CD63 expression. CD63 expression was significantly higher (P = 0.0002) in on-chip activated basophils compared with nonactivated cells. The difference in CD63 expression on anti-FceRI-activated captured basophils in microfluidic chip was significantly higher (P = 0.03) in patients with allergies compared with healthy controls, and the results were comparable with flow cytometry analysis (P = 0.04). Furthermore, there was no significant difference of CD63% expression in anti-FceRI-activated captured basophils in microfluidic chip compared with flow cytometry. Conclusions: We report on the miBAT. This device is capable of isolating basophils directly from whole blood for on-chip activation and detection. The new miBAT method awaits validation in larger patient populations to assess performance in diagnosis and monitoring of patients with allergies at the point of care.
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| 2. |
- Bransky, Avishay, et al.
(författare)
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A Novel Approach to Hematology Testing at the Point of Care
- 2021
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Ingår i: The journal of applied laboratory medicine. - : Oxford University Press (OUP). - 2576-9456 .- 2475-7241. ; 6:2, s. 532-542
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Tidskriftsartikel (refereegranskat)abstract
- BACKGROUND: The need for rapid point-of-care (POC) diagnostics is now becoming more evident due to the increasing need for timely results and improvement in healthcare service. With the recent COVID-19 pandemic outbreak, POC has become critical in managing the spread of disease. Applicable diagnostics should be readily deployable, easy to use, portable, and accurate so that they fit mobile laboratories, pop-up treatment centers, field hospitals, secluded wards within hospitals, or remote regions, and can be operated by staff with minimal training. Complete blood count (CBC), however, has not been available at the POC in a simple-to-use device until recently. The HemoScreen, which was recently cleared by the FDA for POC use, is a miniature, easy-to-use instrument that uses disposable cartridges and may fill this gap.CONTENT: The HemoScreen's analysis method, in contrast to standard laboratory analyzers, is based on machine vision (image-based analysis) and artificial intelligence (AI). We discuss the different methods currently used and compare their results to the vision-based one. The HemoScreen is found to correlate well to laser and impedance-based methods while emphasis is given to mean cell volume (MCV), mean cell hemoglobin (MCH), and platelets (PLT) that demonstrate better correlation when the vision-based method is compared to itself due to the essential differences between the underlying technologies.SUMMARY: The HemoScreen analyzer demonstrates lab equivalent performance, tested at different clinical settings and sample characteristics, and might outperform standard techniques in the presence of certain interferences. This new approach to hematology testing has great potential to improve quality of care in a variety of settings.
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| 3. |
- Isaksson, Anders, et al.
(författare)
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High-Throughput LC-MS/MS Method for Determination of the Alcohol Use Biomarker Phosphatidylethanol in Clinical Samples by Use of a Simple Automated Extraction Procedure-Preanalytical and Analytical Conditions
- 2018
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Ingår i: The Journal of Applied Laboratory Medicine. - : Oxford University Press (OUP). - 2576-9456 .- 2475-7241. ; 2:6, s. 880-892
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Tidskriftsartikel (refereegranskat)abstract
- BACKGROUND: Phosphatidylethanol (PEth) is an alcohol use biomarker with higher clinical sensitivity and specificity than commonly used alcohol markers. Since its introduction as a clinical alcohol-marker in 2006, the number of samples sent to our laboratory for the determination of PEth has shown a strong annual increase. This has prompted the need to develop a cost-effective and reliable analytical procedure with high capacity. METHODS: An LC-MS/MS method for the determination of PEth 16:0/18:1 with a short turnaround time (3 min) has been evaluated with respect to accuracy, sensitivity, and precision. We compared this method with a previously used HPLC method, as well as a manual and a simplified automated method for sample workup, and investigated potential causes of analytic and preanalytic errors. RESULTS: The method shows limits of detection and quantification of 0.0075 μmol/L (5.2 ng/mL) and <0.05 μmol/L (<35 ng/mL), respectively. During a 2.1-year period, the method has shown a total CV < 8% for control samples (n = 2808) in the range of 0.10 (70) to 3.5 μmol/L (2461 ng/mL). The simplified automated method for sample preparation works equally well as the manual one. No specific and clinically significant causes of preanalytic errors were found. CONCLUSIONS: This LC-MS/MS method with automated sample workup is well suited for a clinical laboratory with LC-MS/MS experience and has the capability, proven from several years of use, to produce reliable PEth results in a high-volume laboratory (>50000 clinical samples/year).
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| 4. |
- Lethin, Kajsa, et al.
(författare)
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Effects of 12 Months' Treatment with Testosterone Undecanoate on Markers for Erythropoietic Activity and Safety Aspects in Transgender and Cisgender Hypogonadal Men
- 2023
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Ingår i: The Journal of Applied Laboratory Medicine. - : OXFORD UNIV PRESS INC. - 2576-9456 .- 2475-7241.
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Tidskriftsartikel (refereegranskat)abstract
- Background To investigate the erythropoietic activity and safety aspects of testosterone undecanoate (TU) injections in transgender men, assigned female at birth.Methods Twenty-three men (13 hypogonadal cisgender men and 10 transgender men) who initiated TU at the study start (naive) and 15 men (10 hypogonadal cisgender men and 5 transgender men) on steady-state treatment with TU (non-naive) were included in this prospective 1-year observational study. A control group of 32 eugonadal cisgender men was investigated once at baseline. Complete blood count, testosterone in serum and saliva, and plasma lipids, and liver enzymes were assessed.Results For naive transgender men, a significant increase in hemoglobin concentration was noted (mean (SD)), 141 (8) g/L to 151 (13) g/L, while no increase was seen in naive hypogonadal cisgender men. At the end of the study, naive transgender men exhibited comparable levels of hemoglobin, hematocrit, and testosterone levels in serum and saliva to hypogonadal cisgender men, as well as to the eugonadal cisgender men. During the study, HDL-cholesterol decreased significantly in naive transgender men, 1.4 (0.4) mmol/L to 1.2 (0.4) mmol/L, P = 0.03, whereas no significant change was noted in naive hypogonadal cisgender men. Liver enzymes remained unchanged in all groups.Conclusions After 12 months of treatment with TU in naive transgender men, hemoglobin and hematocrit increased to levels within the cisgender male reference range. A slight decrease in HDL-cholesterol was seen in naive transgender men but liver enzymes remained unchanged.
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| 5. |
- Löfgren, Lars, et al.
(författare)
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Patient-Centric Quantitative Microsampling for Accurate Determination of Urine Albumin to Creatinine Ratio (UACR) in a Clinical Setting
- 2024
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Ingår i: The Journal of Applied Laboratory Medicine. - : Oxford University Press (OUP). - 2576-9456 .- 2475-7241. ; 9:2, s. 329-341
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Tidskriftsartikel (refereegranskat)abstract
- Background: Developing and implementing new patient-centric strategies for drug trials lowers the barrier to participation for some patients by reducing the need to travel to research sites. In early chronic kidney disease (CKD) trials, albuminuria is the key measure for determining treatment effect prior to pivotal kidney outcome trials. Methods: To facilitate albuminuria sample collection outside of a clinical research site, we developed 2 quantitative microsampling methods to determine the urinary albumin to creatinine ratio (UACR). Readout was performed by LC-MS/MS. Results: For the Mitra device the within-batch precision (CV%) was 2.8% to 4.6% and the between-batch precision was 5.3% to 6.1%. Corresponding data for the Capitainer device were 4.0% to 8.6% and 6.7% to 9.0%, respectively. The storage stability at room temperature for 3 weeks was 98% to 103% for both devices. The recovery for the Mitra and Capitainer devices was 104% (SD 7.0%) and 95 (SD 7.4%), respectively. The inter-assay comparison of UACR assessment generated results that were indistinguishable regardless of microsampling technique. The accuracy based on LC-MS/MS vs analysis of neat urine using a clinical chemistry analyzer was assessed in a clinical setting, resulting in 102 ± 8.0% for the Mitra device and 95 ± 10.0% for the Capitainer device. Conclusions: Both UACR microsampling measurements exhibit excellent accuracy and precision compared to a clinical chemistry analyzer using neat urine. We applied our patient-centric sampling strategy to subjects with heart failure in a clinical setting. Precise UACR measurements using quantitative microsampling at home would be beneficial in clinical drug development for kidney therapies.
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| 6. |
- Rehfeld, Linda, et al.
(författare)
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Novel Methods for the Quantification of Dipeptidyl Peptidase 3 (DPP3) Concentration and Activity in Human Blood Samples
- 2019
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Ingår i: The Journal of Applied Laboratory Medicine. - : Oxford University Press (OUP). - 2576-9456 .- 2475-7241. ; 3:6, s. 943-953
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Tidskriftsartikel (refereegranskat)abstract
- BACKGROUND: The ubiquitously expressed dipeptidyl peptidase 3 (DPP3) is involved in protein metabolism, blood pressure regulation, and pain modulation. These diverse functions of DPP3 are attributed to the degradation of bioactive peptides like angiotensin II. However, because of limitations in currently available assays for determination of active DPP3 in plasma, the exact physiological function of DPP3 and its role in the catabolism of bioactive peptides is understudied. Here, we developed 2 assays to specifically detect and quantify DPP3 protein and activity in plasma and validated DPP3 quantification in samples from critically ill patients. METHODS: Assay performance was evaluated in a sandwich-type luminometric immunoassay (LIA) and an enzyme capture activity assay (ECA). DPP3 plasma concentrations and activities were detected in a healthy, population-based cohort and in critically ill patients suffering from severe sepsis and septic shock. RESULTS: The DPP3-LIA and DPP3-ECA show an almost ideal correlation and very similar and robust performance characteristics. DPP3 activity is detectable in plasma of predominantly healthy subjects with a mean (±SD) of 58.6 (±20.5) U/L. Septic patients show significantly increased DPP3 plasma activity at hospital admission. DPP3 activities further increase in patients with more severe conditions and high mortality risk. CONCLUSION: We developed 2 highly specific assays for the detection of DPP3 in plasma. These assays allow the use of DPP3 as a biomarker for the severity of acute clinical conditions and will be of great value for future investigations of DPP3's role in bioactive peptide degradation in general and the angiotensin II pathway in specific.
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| 7. |
- Venge, Per, et al.
(författare)
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Diagnosis and Monitoring of Acute Infections with Emphasis on the Novel Biomarker Human Neutrophil Lipocalin
- 2019
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Ingår i: The Journal of Applied Laboratory Medicine. - : AMER ASSOC CLINICAL CHEMISTRY. - 2576-9456 .- 2475-7241. ; 3:4, s. 664-674
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Forskningsöversikt (refereegranskat)abstract
- Background: Acute infections affect all of us at least once or twice a year. Sometimes the infection prompts a visit to our doctor, and the question asked by the patient and the doctor is whether the infection should be treated with antibiotics or not. This is an important question because unnecessary prescription of antibiotics adds to the increasing problem of antibiotics resistance. Objective means to determine whether the infection is caused by bacteria or virus, therefore, are necessary tools for the doctor. Content: White blood cell counts, C-reactive protein, and other acute-phase reactants in blood are important tools and are commonly used, but unfortunately lack in sensitivity and specificity. In this review we describe some novel biomarkers with increased clinical performance in this regard. The superior biomarker is human neutrophil lipocalin (HNL), a protein released from activated blood neutrophils. HNL may be measured in serum, plasma, or in whole blood after activation with a neutrophil activator. The diagnostic accuracy in the distinction between bacterial and viral acute infections was shown to be in the range of 90%-95% when measured in serum or activated whole blood. Summary: A point-of-care assay for the measurement of HNL in whole blood is currently being developed, which will allow the diagnosis of acute infections within 5-10 min. For certain indications, HNL measurement may be complemented by 1 or 2 other biomarkers, which may increase the diagnostic discrimination between bacterial and viral infections even further.
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