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制定海龟血栓弹力图检测方案。

Establishing a protocol for thromboelastography in sea turtles.

作者信息

Barratclough Ashley, Hanel Rita, Stacy Nicole I, Ruterbories Laura K, Christiansen Emily, Harms Craig A

机构信息

Department of Large Animal Clinical Sciences, University of Florida, College of Veterinary Medicine, Gainesville, Florida, USA.

Department of Clinical Sciences, College of Veterinary Medicine, North Carolina State University, Raleigh, North Carolina, USA.

出版信息

Vet Rec Open. 2018 Aug 13;5(1):e000240. doi: 10.1136/vetreco-2017-000240. eCollection 2018.

Abstract

Thromboelastography (TEG) provides a global evaluation of haemostasis. This diagnostic test is widely used in mammals but has not previously been performed in reptiles, mainly due to the limited availability of taxon-specific reagents. The objective of this pilot study was to establish a protocol to perform TEG in sea turtles. Pooled citrated plasma, stored at -80°C, from four green turtles () was assayed on a TEG 5000. Several initiators were evaluated: kaolin (n=2), RapidTEG (n=2), fresh (n=2) and frozen (n=6) thromboplastin extracted from pooled brain tissue from several chelonian species, human recombinant tissue factor at 1:100 (n=1), Reptilase (n=2), and rabbit thromboplastin (n=1). Both fresh and frozen chelonian thromboplastin were superior in producing quantifiable TEG reaction time compared with all other reagents. These findings are consistent with the lack of an intrinsic pathway in turtles and confirmed a lack of coagulation in the turtle samples in response to mammalian thromboplastin. A TEG protocol was subsequently established for harvested species-specific frozen thromboplastin. The frozen thromboplastin reagent remained stable after one year of storage at -80°C. The developed protocol will be useful as a basis for future studies that aim to understand the pathophysiology of haemostatic disorders in various stranding conditions of sea turtles.

摘要

血栓弹力图(TEG)可对止血情况进行全面评估。这项诊断测试在哺乳动物中广泛应用,但此前尚未在爬行动物中开展,主要是因为特定分类群试剂的可用性有限。这项初步研究的目的是建立在海龟中进行TEG检测的方案。对四只绿海龟保存在-80°C的枸橼酸盐血浆混合样本在血栓弹力图5000仪上进行检测。评估了几种启动剂:高岭土(n = 2)、快速TEG(n = 2)、从几种龟类物种的脑组织中提取的新鲜(n = 2)和冷冻(n = 6)凝血活酶、1:100的人重组组织因子(n = 1)、蛇毒凝血酶(n = 2)以及兔凝血活酶(n = 1)。与所有其他试剂相比,新鲜和冷冻的龟类凝血活酶在产生可量化的TEG反应时间方面更具优势。这些发现与海龟缺乏内源性凝血途径一致,并证实了海龟样本对哺乳动物凝血活酶无凝血反应。随后为收获的物种特异性冷冻凝血活酶建立了TEG检测方案。冷冻凝血活酶试剂在-80°C储存一年后仍保持稳定。所制定的方案将作为未来研究的基础,这些研究旨在了解海龟在各种搁浅情况下止血障碍的病理生理学。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/0ed4/6109949/895b88252b9e/vetreco-2017-000240f01.jpg

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