Stephanie G Crawford, Robert H Coker, Lorrie D Rea
{"title":"使用斯特勒海狮幼崽的全血和血浆,对护理点血酮计和参考方法进行初步比较。","authors":"Stephanie G Crawford, Robert H Coker, Lorrie D Rea","doi":"10.1093/conphys/coad104","DOIUrl":null,"url":null,"abstract":"<p><p>We evaluated the Precision Xtra™ ketometer as part of a larger study categorizing fasting status of free-ranging Steller sea lion (<i>Eumetopias jubatus</i>; SSL) pups which necessitated the identification of plasma β-hydroxybutyrate concentrations ([β-HBA]) around a threshold of <0.3 and ≥0.3 mmol/l. Whole blood samples mixed with sodium heparin (NaHep) or ethylenediaminetetraacetic acid liquid anticoagulants were tested <10 minutes after collection (<i>n</i> = 14; triplicate technical replicates). Plasma (stored at -80°C, NaHep, <i>Thaw1</i>) measured via our laboratory's <i>Reference Assay</i> (Sigma Aldrich, St. Louis, MO, Kit #MAK041) served as the standard [β-HBA] for ketometer comparisons. Our observed β-HBA range (0.0-1.6 mmol/l), consistent with published [β-HBA] of free-ranging Otariid pups, represented the lower 20% of the ketometer's range (0.0-8.0 mmol/l). The maximal coefficient of variation (%CV) of ketometer technical replicates was 9.1% (NaHep, whole blood). The majority of ketometer technical replicate sets (84%, including all matrices, anticoagulants and thawings) were identical (CV = 0%). We found linear relationships and agreement of ketometer [β-HBA] between whole blood preserved with different anticoagulants and between whole blood and plasma (<i>Thaw1</i>) measurements. The ketometer produced results with linearity to the <i>Reference Assay</i> for both whole blood and plasma (<i>Thaw1</i>). We identified a non-linear relationship between plasma at <i>Thaw1</i> and <i>Thaw2</i> (tested four months apart, NaHep), as only samples with higher SSL [β-HBA] decreased in concentration, and all others remained the same. With respect to categorizing SSL pup fasting in our larger study, the ketometer's <i>%Accuracy</i>, %<i>Sensitivity</i> and %<i>Specificity</i> for samples with <i>Reference Assay</i> β-HBA <0.2 and >0.4 mmol/l were 100%. We adopted a modified procedure: plasma samples with mean ketometer concentrations ±0.1 mmol/l of 0.3 mmol/l β-HBA were re-evaluated using the <i>Reference Assay</i>, improving measurement precision from tenths (ketometer) to thousandths (assay) mmol/l. The Precision Xtra™ ketometer was valuable to our application over the range of [β-HBA] observed in SSL pup plasma and whole blood samples.</p>","PeriodicalId":54331,"journal":{"name":"Conservation Physiology","volume":null,"pages":null},"PeriodicalIF":2.6000,"publicationDate":"2024-01-27","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10823332/pdf/","citationCount":"0","resultStr":"{\"title\":\"Preliminary comparisons between a point-of-care ketometer and reference method using Steller sea lion pup whole blood and plasma.\",\"authors\":\"Stephanie G Crawford, Robert H Coker, Lorrie D Rea\",\"doi\":\"10.1093/conphys/coad104\",\"DOIUrl\":null,\"url\":null,\"abstract\":\"<p><p>We evaluated the Precision Xtra™ ketometer as part of a larger study categorizing fasting status of free-ranging Steller sea lion (<i>Eumetopias jubatus</i>; SSL) pups which necessitated the identification of plasma β-hydroxybutyrate concentrations ([β-HBA]) around a threshold of <0.3 and ≥0.3 mmol/l. Whole blood samples mixed with sodium heparin (NaHep) or ethylenediaminetetraacetic acid liquid anticoagulants were tested <10 minutes after collection (<i>n</i> = 14; triplicate technical replicates). Plasma (stored at -80°C, NaHep, <i>Thaw1</i>) measured via our laboratory's <i>Reference Assay</i> (Sigma Aldrich, St. Louis, MO, Kit #MAK041) served as the standard [β-HBA] for ketometer comparisons. Our observed β-HBA range (0.0-1.6 mmol/l), consistent with published [β-HBA] of free-ranging Otariid pups, represented the lower 20% of the ketometer's range (0.0-8.0 mmol/l). The maximal coefficient of variation (%CV) of ketometer technical replicates was 9.1% (NaHep, whole blood). The majority of ketometer technical replicate sets (84%, including all matrices, anticoagulants and thawings) were identical (CV = 0%). We found linear relationships and agreement of ketometer [β-HBA] between whole blood preserved with different anticoagulants and between whole blood and plasma (<i>Thaw1</i>) measurements. The ketometer produced results with linearity to the <i>Reference Assay</i> for both whole blood and plasma (<i>Thaw1</i>). We identified a non-linear relationship between plasma at <i>Thaw1</i> and <i>Thaw2</i> (tested four months apart, NaHep), as only samples with higher SSL [β-HBA] decreased in concentration, and all others remained the same. With respect to categorizing SSL pup fasting in our larger study, the ketometer's <i>%Accuracy</i>, %<i>Sensitivity</i> and %<i>Specificity</i> for samples with <i>Reference Assay</i> β-HBA <0.2 and >0.4 mmol/l were 100%. We adopted a modified procedure: plasma samples with mean ketometer concentrations ±0.1 mmol/l of 0.3 mmol/l β-HBA were re-evaluated using the <i>Reference Assay</i>, improving measurement precision from tenths (ketometer) to thousandths (assay) mmol/l. 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Preliminary comparisons between a point-of-care ketometer and reference method using Steller sea lion pup whole blood and plasma.
We evaluated the Precision Xtra™ ketometer as part of a larger study categorizing fasting status of free-ranging Steller sea lion (Eumetopias jubatus; SSL) pups which necessitated the identification of plasma β-hydroxybutyrate concentrations ([β-HBA]) around a threshold of <0.3 and ≥0.3 mmol/l. Whole blood samples mixed with sodium heparin (NaHep) or ethylenediaminetetraacetic acid liquid anticoagulants were tested <10 minutes after collection (n = 14; triplicate technical replicates). Plasma (stored at -80°C, NaHep, Thaw1) measured via our laboratory's Reference Assay (Sigma Aldrich, St. Louis, MO, Kit #MAK041) served as the standard [β-HBA] for ketometer comparisons. Our observed β-HBA range (0.0-1.6 mmol/l), consistent with published [β-HBA] of free-ranging Otariid pups, represented the lower 20% of the ketometer's range (0.0-8.0 mmol/l). The maximal coefficient of variation (%CV) of ketometer technical replicates was 9.1% (NaHep, whole blood). The majority of ketometer technical replicate sets (84%, including all matrices, anticoagulants and thawings) were identical (CV = 0%). We found linear relationships and agreement of ketometer [β-HBA] between whole blood preserved with different anticoagulants and between whole blood and plasma (Thaw1) measurements. The ketometer produced results with linearity to the Reference Assay for both whole blood and plasma (Thaw1). We identified a non-linear relationship between plasma at Thaw1 and Thaw2 (tested four months apart, NaHep), as only samples with higher SSL [β-HBA] decreased in concentration, and all others remained the same. With respect to categorizing SSL pup fasting in our larger study, the ketometer's %Accuracy, %Sensitivity and %Specificity for samples with Reference Assay β-HBA <0.2 and >0.4 mmol/l were 100%. We adopted a modified procedure: plasma samples with mean ketometer concentrations ±0.1 mmol/l of 0.3 mmol/l β-HBA were re-evaluated using the Reference Assay, improving measurement precision from tenths (ketometer) to thousandths (assay) mmol/l. The Precision Xtra™ ketometer was valuable to our application over the range of [β-HBA] observed in SSL pup plasma and whole blood samples.
期刊介绍:
Conservation Physiology is an online only, fully open access journal published on behalf of the Society for Experimental Biology.
Biodiversity across the globe faces a growing number of threats associated with human activities. Conservation Physiology will publish research on all taxa (microbes, plants and animals) focused on understanding and predicting how organisms, populations, ecosystems and natural resources respond to environmental change and stressors. Physiology is considered in the broadest possible terms to include functional and mechanistic responses at all scales. We also welcome research towards developing and refining strategies to rebuild populations, restore ecosystems, inform conservation policy, and manage living resources. We define conservation physiology broadly and encourage potential authors to contact the editorial team if they have any questions regarding the remit of the journal.