Bench-top Analyzer Research Articles

A-326 Testing for Multidrug-Resistant Acinetobacter baumannii with a Microfluidic Device

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.

B-038 A Novel Ultrasensitive Single-Molecule Counting Technology for Quantitation of Low Abundance Biomarkers

Abstract Background There is a need for sensitive, robust, and scalable analytical methods for accurate, early detection of disease using less invasive sampling methods. Often, smaller volumes and low marker concentrations present challenges to achieving sufficient sensitivity and accuracy with traditional immunoassay methods. This is particularly the case for measuring markers for Alzheimer's and other neurological diseases in blood, where levels are typically at much lower concentrations than in CSF. Here we introduce a novel ultrasensitive technology for flow-based single molecule detection in an optofluidic chip. We have developed an Ultrasensitive Detection Module (UDM) device and optofluidic cartridge that enable easy integration of this technology into existing immunoassay systems. UDM sensitivity data is shown for three prototype biomarker assays in comparison to the Lumipulse G1200 (Fujirebio Inc., Japan). We also present analytical validation data for three Alzheimer’s Disease plasma biomarker assays developed on a fully automated benchtop analyzer with integrated UDM detector. Methods Ultrasensitive detection is achieved by integrated low-loss waveguide in an optofluidic chip that enables maximal excitation of fluorescent molecules traveling through a narrow fluidic channel. The resulting high-intensity emission is detected as a single digital event, enabling individual molecules to be counted without need for amplification. Fluorescence-based immunoassays were developed for detection with the UDM using high performance monoclonal antibodies and reagents (Fujirebio Inc., Japan). Capture antibodies were immobilized on magnetic particles and detection antibodies conjugated to a fluorescent reporter construct (“reporter-Ab”). Multiple incubation and wash steps were followed by dissociation of immune complexes and separation of reporter-Ab from magnetic particles. Released reporter-Ab molecules were injected into the UDM device for digital detection. Signal to noise performance was evaluated in the UDM using reporter construct alone (no assay), as well as for prototype PSA, IL-6, and TSH assays, to establish baseline performance for analytical sensitivity. Plasma assays for pTau181, Aβ1-40, and Aβ1-42 were developed and optimized for the automated benchtop analyzer and validated for analytical sensitivity (LOD/LOQ), linearity, parallelism, precision and repeatability using clinical samples. Results Detection signal-to-noise testing of the reporter construct alone (no immunoassay) demonstrated up to 1000-fold higher sensitivity than the Lumipulse G1200. Prototype assays for PSA, TSH, and IL-6 showed up to 118-fold, 36-fold, and 80-fold higher signal to noise ratio, respectively, when compared to the Lumipulse G1200. Plasma assays for pTau181, Aβ1-40, and Aβ1-42 showed preliminary LLOQs calculated at 0.03 pg/mL, 0.17 pg/mL and 0.37 pg/mL, respectively. Conclusions We have developed a new single molecule counting platform able to detect neurological biomarkers at sub-pg/ml levels in plasma, with the potential to provide sensitive and accurate diagnostic testing of low abundance biomarkers in blood. Such technologies can create new clinical value through higher detection sensitivity and accuracy in areas like Alzheimer’s and other neurological diseases, supporting clinical research and translation to clinical practice.

Combined Lactate Glucose Ratio as a Novel Marker for Rapid Diagnosis of Cerebrospinal Fluid Bacterial Infection in Neurosurgical Patients: Diagnostic Accuracy Study and Benchtop Analyzer Correlation

ObjectiveThis study aimed to assess the diagnostic accuracy of a novel marker, the combined lactate-to-glucose ratio (CLGR), in identifying cerebrospinal fluid (CSF) bacterial infection (CBI) in neurosurgical patients. Additionally, it seeks to establish cut-off values for CLGR and evaluate the reliability of measurement using blood gas analyzer (BGA). MethodsCSF samples were collected from two neurosurgical centers in Kuala Lumpur, Malaysia, between January 2022 and October 2023. Conventional markers and CLGR were quantified using standard laboratory methods, with BGA utilized for measurement when feasible. Samples were categorized into confirmed CBI-positive (CBI+) and CBI-negative (CBI-) groups. Marker performance was compared, and receiver operating characteristic analysis conducted. Pearson's correlation assessed the agreement between BGA and laboratory measurements. ResultsAmong the 130 CSF samples, 11 were CBI+. Both cLac and CLGR were significantly elevated in the CBI+ group (p<0.001). The area under the curve (AUC) for cLac and CLGR was 0.990 and 0.994, respectively. Using a cut-off of 6.0mmol/L, cLac demonstrated sensitivity of 100%, specificity of 93.3%, positive predictive value (PPV) of 57.9%, negative predictive value (NPV) of 100%, and diagnostic accuracy of 93.9%. CLGR ≥20.0 showed even higher accuracy: 100.0% sensitivity, 98.6% specificity, 84.6% PPV, 100% NPV, and overall accuracy of 98.5%. Both markers maintained excellent performance in blood-stained CSF. BGA measurements correlated well with laboratory results (r=0.980 & 0.999, respectively, p<0.001). ConclusionsCLac levels ≥6.0mmol/L and CLGR ≥20.0 accurately identified CBI in neurosurgical patients, with CLGR exhibiting superior efficacy. The potential for instant BGA measurement suggests promise for point-of-care testing.

Comparison of lactate measurements from earlobe and fingertip capillary blood using Biosen S-Line and lactate scout analyzers.

This study aimed to compare variations between the earlobe and fingertip sampling sites in exercises dominated by upper body muscle exertion. It also sought to investigate capillary blood lactate differences between Lactate Scout 4 (LS4) and a bench-top analyzer (Biosen S-Line analyzer, BSL) during Double Poling. Blood samples were collected from the earlobe and fingertip immediately before exercise, at the end of each of five stages, and at 1-, 3-, 5-, and 7-min post-exercise. Forty healthy university students participated as volunteers. During the study, they performed double poling on a ski ergometer with progressively increasing load. Lactate levels were measured using both the BSL and LS4 analyzers. Fingertip Bla values were significantly higher than earlobe values, with a mean bias of -0.66mmol/L, reaching -0.86mmol/L in the 4-8mmol/L range. At the earlobe, the highest CCC between BSL and LS4-a was 0.84 (> 8mmol/L), and for BSL and LS4-b, it was 0.85 (> 8mmol/L). At the fingertip, the highest CCC between BSL and LS4-c was 0.68 (> 8mmol/L), and for BSL and LS4-d, it was 0.52 (> 8mmol/L). Comparing LS4-a and LS4-b at the earlobe, the highest CCC was 0.83 (0-4mmol/L). At the fingertip, comparing LS4-c and LS4-d, the highest CCC was 0.68 (> 8mmol/L). Blood lactate concentrations are higher at the fingertip than the earlobe during SkiErg double poling. The LS4 is less reliable, especially at the fingertip, so using the earlobe with the BSL analyzer is recommended for accurate measurements.

A randomized, controlled trial examining quarter-level somatic cell count and culture-based selective dry cow therapy against blanket dry cow therapy on early lactation production outcomes

The objective of this study was to determine quarters requiring antimicrobial treatment using either a benchtop somatic cell counter or culture with gram-positive selective media and compare the outcomes in these cows to those receiving blanket dry cow therapy (BDCT) in a randomized, controlled trial. We evaluated 2 novel methods of identifying cows with intramammary infections followed by selective antimicrobial treatment at a commercial dairy farm to determine their usefulness in decreasing antibiotic usage during the dry period without significant detrimental effects on milk quality and production. Cows (n = 840) were randomly allocated to one of 3 groups (BDCT, gram-positive selective media culture-based selective dry cow therapy [C-SDCT], and somatic cell count-based SDCT [S-SDCT]) the day before dry-off, and quarter-level milk samples (QLMS) were collected. The QLMS from cows in the S-SDCT group were evaluated using the cell counter, and quarters were treated if SCC was ≥200,000 cells/mL, whereas the QLMS from cows in the C-SDCT group were cultured, and quarters were treated if the culture showed growth. All cows in the BDCT received antimicrobial therapy, and all cows received an internal teat sealant regardless of treatment group. Outcomes measured were first and second DHIA test SCC, milk production through 60 DIM, cows leaving the farm, clinical mastitis, and bacteriologic new infections in a subset of quarters. Cows in both SDCT groups had fewer antimicrobial treatments than cows in the BDCT group as was expected, and cows in the C-SDCT group had fewer treatments than those in the S-SDCT group. Cows in both SDCT groups had a higher linear score at the first DHIA test (BDCT: 1.8, S-SDCT: 2.2, C-SDCT: 2.2); however, we found no other differences between groups regarding any other outcomes measured. Although antimicrobial use was significantly reduced, farms should use caution in adopting the benchtop analyzer and the selective media described in this study as ways to identify infected cows for dry cow therapy because they may result in increased linear scores early in lactation.

Noninvasive and Multiplex Self-Test of Kidney Disease Biomarkers with Graphene-Based Lab-on-a-Chip (G-LOC): Toward Digital Diagnostics in the Hands of Patients.

Chronic kidney disease is one of the major health issues worldwide. However, diagnosis is now highly centralized in large laboratories, resulting in low access to patient monitoring and poor personalized treatments. This work reports the development of a graphene-based lab-on-a-chip (G-LOC) for the digital testing of renal function biomarkers in serum and saliva samples. G-LOC integrates multiple bioelectronic sensors with a microfluidic system that enables multiplex self-testing of urea, potassium, sodium, and chloride. The linearity, limit of detection (LOD), accuracy, and coefficient of variability (CV) were studied. Accuracy values higher than 95.5% and CV lower than 9% were obtained for all of the biomarkers. The analytical performance was compared against three reference lab benchtop analyzers by measuring healthy- and renal-failure-level samples of serum. From receiver operating characteristic (ROC) plots, sensitivities (%) of 99.7, 97.6, 99.1, and 89.0 were obtained for urea, potassium, sodium, and chloride, respectively. Then, the test was evaluated in noninvasive saliva samples and compared against reference methods. Correlation and Bland-Altman plots showed good correlation and agreement of the G-LOC with the reference methods. It is noteworthy that the precision of G-LOC was similar to better than benchtop lab analyzers, with the advantage of being highly portable. Finally, a user testing study was conducted. The analytical performance obtained with untrained volunteers was similar to that obtained with trained chemists. Additionally, based on a user experience survey, G-LOC was found to have very simple usability and would be suitable for at-home diagnostics.

Programmable flow injection: a versatile technique for benchtop and autonomous analysis of phosphate and silicate in seawater

High-resolution, autonomous monitoring of phosphate and silicate in the marine environment is essential to understand their complex dynamics and implications for the functioning of marine ecosystems. In the absence of dependable reagent-less sensors for these nutrients, leveraging established colorimetric techniques using miniaturized analyzers, such as programmable Flow Injection (pFI), offers the best immediate solution to meet oceanographic accuracy and precision standards. In this work, we further optimize the phosphomolybdate and silicomolybdate assays recently adapted for use with pFI, laying the groundwork for the technique’s use for long-term, autonomous operations. For both assays, we show that only a narrow range of acidities and molybdate concentrations can maximize sensitivity while minimizing salt effects. In addition, we demonstrate the stability of our optimized colorimetric reagent formulations, ensuring that analytical sensitivity remains within 10% of initial levels for at least 35 days of continuous use. We then applied our optimized protocols to produce oceanographically consistent phosphate and silicate profiles at the Hawaii Ocean Time Series (HOTS) and Southern Ocean Time Series (SOTS), respectively, which compared favorably against a reference method and historical data. Using certified reference materials for nutrients in seawater, we show that our pFI protocols, optimized for long-term operations, achieve a shipboard precision better than 6% and a relative combined uncertainty (k=1) of 4.5% for phosphate (0.45 - 2.95 µmol L-1) and 6.2% for silicate (2.2 to 103 µmol L-1). To demonstrate pFI’s potential as a versatile tool for autonomous monitoring, we report a five-day hourly phosphate time series at a coastal shore station in central California (n=121 analyses), examine phosphate uptake by seaweed at five-minute intervals at a seaweed aquaculture facility (n=103), and discuss a unique, high-resolution surface silicate transect spanning multiple frontal zones in the Australian sector of the Southern Ocean (n=249). These data, obtained using a commercially available pFI analyzer, confirm that pFI is a viable technology for autonomous monitoring of phosphate and silicate, paving the way for more ambitious, long-term deployments in a variety of settings.

Measuring the Level of Agreement for Lactate Measurements in Hispaniolan Amazon Parrots (Amazona ventralis) Among 2 Point-of-Care Analyzers and a Benchtop Analyzer.

Lactate is an important biochemistry analyte used in human and veterinary medicine to assess tissue perfusion and can be used as a prognostic indicator for certain disease conditions. Whereas lactate is commonly measured using "patient-side" handheld meters, these meters have not been validated for companion avian species. The purpose of this study was to measure the level of agreement between 2 commercially available point-of-care lactate meters and a laboratory benchtop blood analyzer in Hispaniolan Amazon parrots (Amazona ventralis). Blood samples were collected from 20 adult parrots at Louisiana State University by drawing 1.5 mL of blood from the right jugular vein. One drop of whole blood was used for the Lactate Plus analyzer and the remainder of the sample transferred into a lithium heparin microtainer. From the blood in the microtainer, 0.2 mL whole blood was analyzed using the epoc Blood Analysis System, and the remaining sample was centrifuged to obtain plasma that was immediately frozen at -80°C (-112°F) and submitted to the Texas A&M University Clinical Pathology Laboratory for analysis on the VITROS 4500 benchtop analyzer. Bland-Altman agreement plots and Passing-Bablok regression were used to measure the level of agreement between the methods. There was poor agreement between all 3 methods with mean percentage differences in lactate concentrations ≥22% (epoc and Lactate Plus: 33.6% [95% CI: 27-40]; epoc and VITROS 4500: 55% [95% CI:52-58]; VITROS 4500 and Lactate Plus: 22% [95% CI:16-28]). Based on these results, the point-of-care meters tested in this study are not interchangeable, and separate reference intervals were calculated for each method. Blood lactate concentrations may have more utility in tracing lactate trends over time in an individual rather than being able to utilize this information at 1 time point for disease diagnosis and prognosis.

Clinical evaluation of a fully electronic microfluidic white blood cell analyzer.

The White Blood Cell (WBC) count is one of the key parameters signaling the health of the immune system. Abnormal WBC counts often signal a systemic insult to the body such as an underlying infection or an adverse side effect to medication. Typically, the blood collected is sent to a central lab for testing, and results come back within hours, which is often inconvenient and may delay time-sensitive diagnosis or treatment. Here, we present the CytoTracker, a fully electronic, microfluidic based instant WBC analyzer with the potential to be used at point-of-care. The CytoTracker is a lightweight, portable, affordable platform capable of quantifying WBCs within minutes using only 50 μl of blood (approximately one drop of blood). In this study, we clinically evaluated the accuracy and performance of CytoTracker in measuring WBC and granulocyte counts. A total of 210 adult patients were recruited in the study. We validated the CytoTracker against a standard benchtop analyzer (Horiba Point of Care Hematology Analyzer, ABX Micros 60). Linear dynamic ranges of 2.5 k/μl- 35 k/μl and 0.6 k/μl- 26 k/μl were achieved for total WBC count and granulocyte count with correlation coefficients of 0.97 and 0.98. In addition, we verified CytoTracker's capability of identifying abnormal blood counts with above 90% sensitivity and specificity. The promising results of this clinical validation study demonstrate the potential for the use of the CytoTracker as a reliable and accurate point-of-care WBC analyzer.

Estimation of fluoride release and recharge potential among Zirconomer, GC Gold Label Hybrid, Ketac Molar, and Tetric Prime: An in vitro study

Abstract Background: The preventive role of fluoride in caries control is mainly due to the formation of fluorapatite crystals which are more resistant to acid attack. Glass ionomer cement (GIC), its newer modifications, and certain composites in which fluoride has been incorporated show fluoride release. Aim: This research aims to compare the initial fluoride liberated, the fluoride liberated after recharge, and the mean percentage reduction of modified varieties of GICs along with fluoride-releasing composites. Materials and Methods: Forty specimen blocks of the materials to be tested were divided into 5 groups (n = 8), namely Group 1: Zirconomer (SHOFU), Group 2: GC GOLD Label Hybrid (GC), Group 3: Tetric Prime (Ivoclar), Group 4: Ketac Molar (3M), and Group 5: TE-Econom Plus (Ivoclar). The initial fluoride liberated was measured on the 1st day, after 1 week, and after a fortnight, using a benchtop analyzer and total ionic strength adjustment buffer III analyzer. Then, fluoride application was done with fluoridated dentifrice using the hand brush method, and the samples were rechecked for fluoride release at similar time intervals, i.e. 1, 7, and 15 days. Results: Zirconomer showed significantly higher initial fluoride release. The minimum mass of fluoride was released by TE-Econom. A similar pattern was observed for fluoride liberation after recharge, with Zirconomer showing maximum re-release of fluoride and TE Econom showing maximum reduction in fluoride release after 15 days. Conclusion: Zirconomer exhibited high initial fluoride liberation and fluoride liberation after recharge following topical fluoride application. GC Gold Label Hybrid and Ketac Molar showed the least percentage reduction in the biweekly fluoride liberation which was remarkably lower than Zirconomer. Fluoride-releasing composites showed the least amount of initial fluoride liberation and fluoride liberation after recharge.

Verification of a Benchtop Hematology Analyzer with a 5-Part Differential Count: There is Nothing Wrong with Being Small.

The benchtop ADVIA 560 AL hematology analyzer (Siemens Healthineers Tarrytown, NY, USA) offers a small footprint and ease of operation making it suitable for satellite laboratories and intensive care units. A verification study of this analyzer was performed. Between- and intra-run precision, carry-over, linearity, and throughput were evaluated on the ADVIA 560 AL. Accuracy was assessed on 94 patient samples by comparing the results obtained on the ADVIA 560 AL to the results on the reference Sysmex XN1000 analyzer (Sysmex Corporation, Kobe, Japan). The ADVIA 560 AL showed acceptable imprecision on control material and minimal bias in comparison to the XN 1000 on patient samples with a throughput of 60 samples per hour. The percentage carryover was not significant and the linearity was within acceptable limits. The ADVIA 560 AL bench-top analyzer is suitable for acute care centers and satellite laboratories owing to its small footprint, ease of use, and reproducible and accurate results.

Redo Transcatheter Aortic Valve Implantation in the Lotus Mechanically Expanded Transcatheter Heart Valve: Bench-Top Analysis, Clinical Experience, and Procedural Guidance.

Redo transcatheter aortic valve implantation (TAVI) is increasing as patients outlive their transcatheter heart valves (THVs) and present with bioprosthetic valve failure. The Lotus mechanically expanded THV has unique design characteristics, which have specific implications for Redo TAVI. The design features of the Lotus valve and their relevance to Redo TAVI were reviewed. Bench-top analysis of Redo TAVI was performed using different contemporary THVs. Procedural and outcome data were obtained from 10 patients who had undergone Redo TAVI for Lotus bioprosthetic valve failure in 5 centers. Recommendations for performing Redo TAVI in Lotus are made, based on these findings. The Lotus leaflets extend from the frame inflow, with a Neoskirt of only 13 mm, hence a low risk of coronary obstruction during Redo TAVI. The Lotus frame posts prevent full apposition of the Redo prosthesis in the upper part of the frame, while implantation of the Redo THV above the Lotus inflow leads to inadequate apposition of the Lotus leaflets. Inflow-to-inflow positioning is therefore recommended for effective sealing and leaflet pinning. The Lotus locking mechanism prevents overexpansion of the frame, limiting Redo THV oversizing. Redo TAVI was favorable with SAPIEN 3, Evolut, and Navitor THVs on bench-top analysis but not with ACURATE Neo 2 due to the upper crowns and short stent preventing inflow-to-inflow deployment. Case review demonstrated satisfactory outcomes in 10 patients treated with Evolut (n=6), SAPIEN 3 (n=3), and Portico (n=1) valves, with no mortality, major morbidity, or coronary obstruction. Three patients had residual mean gradient ≥20 mm Hg, including 2 of 3 SAPIEN cases. Guidance on procedural planning, valve choice, sizing, and positioning is provided. Redo TAVI in Lotus requires an understanding of unique design characteristics, and adherence to key procedural recommendations, but can be safely and effectively performed with most contemporary valve types.

Comparative Evaluation of Fluoride Release from Four Commercially Available Pediatric Dental Restorative Materials.

The aim of this study was to evaluate the fluoride-releasing abilities of commercially available restorative materials such as-Activa™ BioActive-restorative™ material, Zirconomer (Shofu Inc), Beautifil® II (Shofu Inc), GC Gold Label 9 high strength posterior restorative glass ionomer cement (GIC Corp). A total of 40 disk specimens (10 of each material) were placed into distilled/deionized (DI) water and the fluoride release was measured for 30 days. Fluoride ion measurement was performed at the end of the 1st, 3rd, 7th, 15th, and 30th day under normal atmospheric conditions by fluoride ion selective electrode (F-ISE) (Orion 9609 BNWP, Ionplus SureFlow fluoride electrode, Thermo Scientific, United States of America) coupled to a benchtop analyzer (Hachsen Ion+). All the materials included in the study exhibited fluoride release. Although there were differences in the amounts of fluoride released between Activa™, Zirconomer, and GC Gold Label 9 the mean difference between these three groups was not found to be statistically significant. Beautifil® II showed low amounts of fluoride released at all time intervals. Among the above-compared materials Activa™ and Zirconomer exhibit both improved mechanical properties as well as they have fluoride-releasing ability so can be preferred over conventional glass ionomer restorations. Dhumal RS, Chauhan RS, Patil V, et al. Comparative Evaluation of Fluoride Release from Four Commercially Available Pediatric Dental Restorative Materials. Int J Clin Pediatr Dent 2023;16(S-1):S6-S12.

Verification of the Bühlmann fCAL turbo faecal calprotectin assay on the Binding Site Optilite benchtop analyser

ObjectivesInflammatory bowel disease (IBD) and irritable bowel syndrome (IBS) are increasingly prevalent disorders. Faecal calprotectin is useful in the differential diagnosis of IBD from IBS and monitoring IBD activity. We verified the Bühlmann fCAL turbo faecal calprotectin assay on the Binding Site, Optilite benchtop analyser. DesignAccuracy, precision, lower limit of quantitation (LLoQ), and linearity of the Bühlmann fCAL turbo faecal calprotectin assay on the Binding Site, Optilite benchtop analyser were ascertained. Comparison with the Bühlmann Quantum Blue fCAL extended and DiaSorin, Liaison calprotectin assays were also undertaken. Difference between assays was evaluated using the Wilcoxon signed-rank test and method comparison was undertaken using Spearman’s rank correlation (rs), difference plots and Passing-Bablok regression analyses. ResultsThe fCAL turbo assay was linear between 25 and 10,000 μg/g, and the LLoQ was 25 μg/g. Intra-, and inter-assay imprecision was <5%. There was a good agreement (rs = 0.96) and no significant bias (3%, p = 0.10) present between the fCAL turbo and Quantum Blue extended assays. Between the fCAL turbo and DiaSorin, liaison assays there was a good agreement (rs = 0.97), but a significant bias (53%, p = <0.01) was present. ConclusionsThe fCAL turbo assay performs well on the Binding Site, Optilite benchtop analyser. Calprotectin results are commutable between with Bühlmann fCAL turbo and Quantum Blue fCAL extended assays, but not between Bühlmann and DiaSorin calprotectin assays.

Mammary gland secretions’ conductivity alone or coupled with pH as a novel and accurate predictor of foaling and its association with electrolytes

Accurate prediction of parturition is paramount to providing assisted delivery and preventing foaling-induced complications. Assessment of pH and electrolyte concentrations of mammary gland secretions (MGS) are useful to detect impending parturition. The study's hypothesis was that changes in electrolytes and pH alter the conductivity of MGS, thus conductivity could be a more accurate foaling predictor than pH and electrolytes alone. In experiment 1, periparturient mares (n=23) had MGS assessed daily for pH, conductivity, and electrolytes (Ca2+, Na+, and K+) using bench-top analyzers (LAQUA-Twin D 771, Horiba LTDA, Kyoto Japan for pH and conductivity; and Hitachi 911 Roche Diagnostics, Basel Switzerland for electrolytes) and portable handheld devices. Handheld devices (LAQUA-Twin, Horiba) were also compared with bench top analyzers. Mixed-model analyses revealed significant reductions in pH, conductivity, Na+, and increases in Ca2+, and K+ upon impending parturition. Bland-Altman analyses showed no significant deviation from linearity for all devices. In experiment 2, periparturient mares (n=114) had daily MGS assessed for conductivity, pH, and electrolytes using hand-held devices (Laqua-Twin).Mixed-model analyses showed effects of time (P<0.05), but no effect of group (P>0.05) or interaction (group*time) (P>0.05) for all analytes. Conductivity was strongly associated with pH (r=0.90), and moderately associated with Na+ (r=0.77), Ca2+ (r= -0.60), and K+ (r= -0.70). Analyses of receiver operating characteristic (ROC) were performed to determine the cut offs for foaling in 24 h for all analytes; and then the cut off values were used to calculate the sensitivity, specificity, positive predictive value (PPV), and negative predictive value (NPV). The results were: conductivity for sensitivity, specificity, PPV, and NPV (0.82, 0.80, 0.82, and 0.60), pH (0.80,0.78,0.81, and 0.70), Ca2+(0.92,0.35,0.68, and 0.76), Na+ (0.65,0.28,0.53, and 0.39), and K+ (0.67,0.25,0.50,0.41). The sensitivity, specificity, PPV, and NPV, for conductivity coupled with pH, were 0.84,0.85, 0.81, and 0.90 respectively. In conclusion, the handheld devices tested herein are as accurate as bench-top analyzers for pH, conductivity, and electrolytes. The conductivity of MGS showed similar results to pH to predict of foaling, and the combination of pH and conductivity enhanced the prediction of parturition in mares.

Using seconds-resolved pharmacokinetic datasets to assess pharmacokinetic models encompassing time-varying physiology.

Pharmacokinetics have historically been assessed using drug concentration data obtained via blood draws and bench-top analysis. The cumbersome nature of these typically constrains studies to at most a dozen concentration measurements per dosing event. This, in turn, limits our statistical power in the detection of hours-scale, time-varying physiological processes. Given the recent advent of in vivo electrochemical aptamer-based (EAB) sensors, however, we can now obtain hundreds of concentration measurements per administration. Our aim in this paper was to assess the ability of these time-dense datasets to describe time-varying pharmacokinetic models with good statistical significance. We used seconds-resolved measurements of plasma tobramycin concentrations in rats to statistically compare traditional one- and two-compartmental pharmacokinetic models to new models in which the proportional relationship between a drug's plasma concentration and its elimination rate varies in response to changing kidney function. We found that a modified one-compartment model in which the proportionality between the plasma concentration of tobramycin and its elimination rate falls reciprocally with time either meets or is preferred over the standard two-compartment pharmacokinetic model for half of the datasets characterized. When we reduced the impact of the drug's rapid distribution phase on the model, this one-compartment, time-varying model was statistically preferred over the standard one-compartment model for 80% of our datasets. Our results highlight both the impact that simple physiological changes (such as varying kidney function) can have on drug pharmacokinetics and the ability of high-time resolution EAB sensor measurements to identify such impacts.

Successful use of a benchtop fluorescent enzyme immunoassay analyzer to measure serum cortisol concentration as a screening test for hypoadrenocorticism in dogs.

To assess the diagnostic performance of a benchtop fluorescent enzyme immunoassay analyzer (AIA-360; Tosoh Bioscience Inc) for the measurement of serum cortisol concentration as a screening test for hypoadrenocorticism in dogs. 173 client-owned dogs (20 with hypoadrenocorticism and 153 with nonadrenal illness). Medical records of all dogs that underwent an ACTH stimulation test between June 2015 and October 2019 were reviewed retrospectively. Dogs were excluded if the ACTH stimulation test was performed on the basis of a suspicion of hypercortisolism, serum cortisol concentrations were measured using an analyzer other than the one assessed in the present study, or dogs had received medication known to affect the pituitary-adrenal axis in the 4 weeks1,2 preceding ACTH stimulation testing. The diagnostic performance of the benchtop analyzer was evaluated by calculating sensitivity, specificity, and likelihood ratios at various cutoff points. Serum resting cortisol cutoff point concentrations of 0.8 μg/dL (22 nmol/L), 1 μg/dL (28 nmol/L), and 2 μg/dL (55 nmol/L) had a sensitivity of 100%. An optimal serum resting cortisol cutoff point of 0.58 μg/dL (16 nmol/L) had a sensitivity, specificity, and positive and negative likelihood ratios of 100%, 97%, and 30.6 and 0.0, respectively. Findings indicated that previously derived cutoff points could be used with excellent sensitivity to exclude hypoadrenocorticism in this population of dogs when serum cortisol concentration was measured with the evaluated benchtop analyzer. An ACTH stimulation test may need to only be performed to diagnose hypoadrenocorticism if resting serum cortisol concentration is ≤ 0.58 μg/dL when measured with the evaluated benchtop analyzer.

Verification of the Bühlmann fPELA turbo faecal elastase assay on the Binding Site Optilite benchtop analyser.

Pancreatic elastase-1 (PE1) can be measured to assess exocrine activity of the pancreas. A semi-automated particle-enhanced, open-channel turbidimetric immunoassay has been introduced by Bühlmann (fPELA turbo, Bühlmann Laboratories AG, Schoenenbuch, Switzerland). Published evaluation data is lacking. We therefore verified performance of the assay on the Binding Site Optilite benchtop analyser and undertook a sample comparison with the DiaSorin PE1 assay on the Liaison. Accuracy, imprecision, lower limit of quantitation (LLoQ) and linearity of the Bühlmann fPELA turbo assay on the Binding Site Optilite analyser was ascertained. Comparison with the DiaSorin Liaison PE1 assay was also undertaken. Difference between assays was evaluated using the Wilcoxon signed-rank test and method comparison was undertaken using Spearman's rank correlation (rs), Bland-Altman and Passing-Bablok regression analyses. The fPELA turbo assay was linear between 5 and 2500μg/g. The LLoQ was 5µg/g. Intra- and inter-assay imprecision was <6%. There was a good agreement (rs = 0.92) and no significant bias (5.8µg/g, P = 0.29) present between the Bühlmann fPELA turbo and DiaSorin PE1 assays. The Bühlmann fPELA turbo assay performs well on the Binding Site Optilite analyser. Faecal elastase results are commutable between with Bühlmann fPELA turbo and DiaSorin Liaison PE1 assays.

Is staple line reinforcement still needed on contemporary staplers? A benchtop analysis.

Staple line reinforcement (SLR) is commonly used in bariatric surgeries to reduce leaks and bleeds. With the evolution of staplers, the need for buttressing with the latest surgical stapling technology is in question. The efficacy of GORE® SEAMGUARD® (G-SLR) to improve staple line strength based on an established measure of burst pressure was evaluated. A benchtop test on synthetic tissue evaluated the pressure required for staple line leak across surgical staplers with and without G-SLR. Staple lines on a consistent thickness synthetic bowel were pressurized to the point of failure (burst pressure) among Ethicon®, Intuitive®, and Medtronic® surgical staplers with and without G-SLR. Burst pressure and leak location (through the staple line [TTSL] or through the staple [TTS], on the anvil or cartridge side) were recorded. Visual confirmation of a leak concluded each test. The pooled mean burst pressure for G-SLR was greater (p < 0.05) by 0.494 pounds/square inch compared with no reinforcement with no meaningful differences among staplers. Leak failures were primarily TTS (91.7%) and equally distributed between reinforcement groups with more leak failures on the cartridge side with G-SLR and on the anvil side for non-SLR group. Leaks occurred across the length of staple lines with no discernable pattern. Employing a buttressing material strengthens the staple line, as measured by burst pressure, and may reduce the risk for staple line failure. This benchtop study of G-SLR with three commonly used surgical staplers demonstrated a significant increase in burst pressures among the studied stapling devices.

Application of the Evidence MultiSTAT benchtop analyser to the fast (under 30 minutes), simultaneous, semi-quantitative determination of 29 drugs of abuse from one urine sample

Biochip array technology allows for simultaneous screening of multiple drugs of abuse from a single human specimen and applied to the fully automated benchtop analyser Evidence MultiSTAT, provides rapid test results. Current technology compares the signal output of a sample to that of a cut-off, reporting a qualitative test result. Further advancement of the Evidence MultiSTAT technology allows a semi-quantitative result to be generated. Embedded software, preloaded with multiple calibration curves for each drug target, selects an optimal calibration curve by running two adjuster levels. Additionally, the software allows a user defined cut-off concentration to be selected enabling two samples to be analysed in parallel in under 30 minutes. The MultiSTAT DOA ToxPlex Urine array reports semi-quantitative test results for 29 drug targets simultaneously from a single sample. Simultaneous competitive chemiluminescent immunoassays ( n = 29) were employed. The assays were applied to the Evidence MultiSTAT biochip analyser, which processes a self-contained cartridge comprising all the components required for the assays. The system software selects the optimal calibration for each drug target from the database and processes and reports the multiple semi-quantitative results generated. The Graphical User Interface (GUI) presents the sample concentration for the 29 drug targets, the user defined cut-off concentration and whether the sample is classified as positive or negative in comparison to the selected cut-off. Inter-Assay Precision and Recovery performance were evaluated; three samples (−50%, cut-off and +50% samples) were assayed in triplicate per run for a total of 5 runs, n = 15. Resulting CVs were < 20% and recovery to target was within 70–130% for all 29 assays, confirming the suitability of the new semi-quantitative functionality. Application of the fully automated Evidence MultiSTAT allows for fast (under 30 minutes) semi-quantitative determination of 29 drug targets. The multiple results generated are displayed in an easy to visualise tabulated format and the system has the capacity to store and archive the results produced, which greatly aids accurate drug screening.