Measurement of plasma cell-free DNA concentrations in dogs with sepsis, trauma, and neoplasia
Measurement of plasma cell-free DNA concentrations in dogs with sepsis, trauma, and neoplasia
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DOI：
10.1111/vec.12592
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发表时间：
2017-05-01
影响因子：
1.3
通讯作者：
Goggs, Robert
中科院分区：
文献类型：
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作者：
Letendre, Jo-Annie;Goggs, Robert
目的 - 确定犬血浆中是否可以识别无细胞 DNA (cfDNA)，评估 3 种测量送至紧急服务机构的犬血浆 cfDNA 浓度的技术，并将健康犬与脓毒症、创伤和肿瘤患者的血浆 cfDNA 浓度进行比较。设计 - 对 2014 年 5 月至 2014 年 12 月期间收集的储存犬血浆样本进行回顾性研究。设置 - 送至大学兽医紧急服务处的犬动物 - 对从 15 只患有败血症的狗、15 只患有中重度创伤的狗和 15 只诊断患有肉瘤的狗获得的残留血浆样本进行血浆 cfDNA 测量。还测量了 15 只健康狗的血浆 cfDNA。干预措施 - 无。测量和主要结果 - 评估了测定线性、重复性和再现性。在稀释的柠檬酸盐血浆上重复进行 cfDNA 定量，并使用 2 种荧光测定（SYBR-Gold；Quant-iT）和紫外吸收光谱进行 DNA 纯化。使用标准曲线将荧光强度 (FI) 转换为 cfDNA 浓度。使用 Kruskal-Wallis 检验将中位 FI 值和 cfDNA 浓度与健康对照进行比较，并进行多重比较调整。 Alpha 设置为 0.05。两种测定均具有出色的线性以及可接受的重复性和再现性。与对照组相比，两种检测中脓毒症或中重度创伤犬的血浆 cfDNA 浓度均显着增加（P
Objectives - To determine if cell-free DNA (cfDNA) was identifiable in canine plasma, to evaluate 3 techniques for the measurement of plasma cfDNA concentrations in dogs presented to an emergency service, and to compare the plasma cfDNA concentrations of healthy dogs to those with sepsis, trauma, and neoplasia.Design - Retrospective study of banked canine plasma samples collected between May 2014 and December 2014.Setting - Dogs presented to the emergency service of a university veterinary teaching hospital.Animals - Plasma cfDNA was measured on residual plasma samples obtained from 15 dogs with sepsis, 15 dogs with moderate-severe trauma, 15 dogs diagnosed with a sarcoma. Plasma cfDNA was also measured in 15 healthy dogs.Interventions - None.Measurements and Main Results - Assay linearity, repeatability, and reproducibility were evaluated. Quantification of cfDNA was performed in duplicate on diluted citrated plasma and following DNA purification using 2 fluorescence assays (SYBR-Gold; Quant-iT) and by ultraviolet absorbance spectroscopy. Fluorescence intensities (FIs) were converted to cfDNA concentrations using standard curves. Median FI values and cfDNA concentrations were compared to healthy controls using the Kruskal-Wallis test, with adjustment for multiple comparisons. Alpha was set at 0.05. Both assays had excellent linearity, and acceptable repeatability and reproducibility. Compared to controls, plasma cfDNA concentrations were significantly increased in dogs with sepsis or moderate-severe trauma with both assays (P