Effects of EDTA on chemiluminescent immunoassay measurement of ACTH, cortisol, and thyroid hormones in dogs

IF 1.9 2区 农林科学 Q2 AGRICULTURE, DAIRY & ANIMAL SCIENCE
A.L. Hirsch , J.M. Fletcher , M.A. Mitchell
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引用次数: 1

Abstract

When measuring blood hormones, pre-analytical sample handling can impact the quality of the results. Previous studies have shown improved stability of canine cortisol in ethylenediaminetetraacetic acid (EDTA) plasma compared to serum and interchangeability of serum and plasma when cortisol is measured by radioimmunoassay. Additionally, cortisol samples were also interchangeable when measured by chemiluminescent immunoassay if the EDTA concentration was consistent with that of optimally filled tubes, whereas excess EDTA interfered with the measurement of cortisol and serum and EDTA plasma were not interchangeable when measuring total thyroxine (TT4). The main limitation of these studies was that they were performed by spiking pooled serum samples with EDTA or in previously collected blood samples submitted to a clinical pathology laboratory. The purpose of the present study was to evaluate the effect of EDTA on the measurement of adrenocorticotropic hormone (ACTH), cortisol, TT4, free thyroxine (FT4), and thyroid stimulating hormone (TSH) in healthy dogs using the Siemens IMMULITE 1000. Whole blood from forty dogs was aliquoted into three Monoject sample tubes: no additive, completely filled EDTA tube, and 50% filled EDTA tube. Handling and storage conditions were identical, and all samples were analyzed on the same day. Bland-Altman plots and Passing-Bablok regression were used to assess agreement and risks for error, respectively. Proportional errors were found between serum and plasma samples for ACTH, cortisol, TT4, FT4, and TSH; systematic errors were also found for FT4. There was poor agreement and clinically significant differences between the measured concentrations of all hormones in serum and plasma, proving that these sample types are not interchangeable. Incompletely filled EDTA tubes were associated with significantly lower ACTH concentrations compared to completely filled EDTA tubes. When measured by chemiluminescent immunoassays that utilize alkaline phosphatase at the reporter enzyme, serum should be used for cortisol, TT4, FT4, and TSH, while plasma from completely filled EDTA tubes should be used for ACTH.

EDTA对狗化学发光免疫测定ACTH、皮质醇和甲状腺激素的影响
当测量血液激素时,分析前的样品处理会影响结果的质量。先前的研究表明,与血清相比,犬乙二胺四乙酸(EDTA)血浆中皮质醇的稳定性更好,并且当用放射免疫法测量皮质醇时,血清和血浆的互换性更好。此外,如果EDTA浓度与最佳填充管的浓度一致,则化学发光免疫分析法测量皮质醇样品时也可以互换,而过量的EDTA会干扰皮质醇的测量,并且在测量总甲状腺素(TT4)时血清和EDTA血浆不能互换。这些研究的主要局限性是,它们是通过将EDTA混合的血清样本或先前收集的血液样本提交给临床病理实验室进行的。本研究的目的是评价EDTA对健康犬促肾上腺皮质激素(ACTH)、皮质醇、TT4、游离甲状腺素(FT4)和促甲状腺激素(TSH)测量的影响。将40只狗的全血分别放入三个Monoject样管中:无添加剂、完全填充EDTA管和填充50% EDTA管。处理和储存条件相同,所有样品在同一天进行分析。Bland-Altman图和Passing-Bablok回归分别用于评估一致性和错误风险。血清和血浆ACTH、皮质醇、TT4、FT4和TSH存在比例误差;FT4也发现了系统误差。在血清和血浆中所有激素的测量浓度之间存在较差的一致性和临床显着差异,证明这些样品类型不可互换。与完全填充的EDTA管相比,未完全填充的EDTA管ACTH浓度显著降低。当使用碱性磷酸酶作为报告酶的化学发光免疫测定时,血清应用于皮质醇、TT4、FT4和TSH,而完全填充的EDTA管中的血浆应用于ACTH。
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来源期刊
Domestic animal endocrinology
Domestic animal endocrinology 农林科学-奶制品与动物科学
CiteScore
5.50
自引率
4.80%
发文量
58
审稿时长
31 days
期刊介绍: Domestic Animal Endocrinology publishes scientific papers dealing with the study of the endocrine physiology of domestic animal species. Those manuscripts utilizing other species as models for clinical or production problems associated with domestic animals are also welcome. Topics covered include: Classical and reproductive endocrinology- Clinical and applied endocrinology- Regulation of hormone secretion- Hormone action- Molecular biology- Cytokines- Growth factors
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