Validation of commercial ELISAs for quantifying anabolic growth factors and cytokines in canine ACD-A anticoagulated plasma
Validation of commercial ELISAs for quantifying anabolic growth factors and cytokines in canine ACD-A anticoagulated plasma
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DOI：
10.1177/1040638717690186
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发表时间：
2017-03-01
影响因子：
1.5
通讯作者：
Franklin, Samuel P.
中科院分区：
文献类型：
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作者：
Birdwhistell, Kate;Basinger, Robert;Franklin, Samuel P.
富含血小板的血浆已在狗身上进行了广泛的研究，但尚未对用柠檬酸盐抗凝剂制备的犬血浆中的合成代谢生长因子和炎症细胞因子进行定量的酶联免疫吸附测定（ELISA）进行验证。我们对转化生长因子-β 1 (TGF-1)、血小板衍生生长因子-BB (PDGF-BB)、血管内皮生长因子 (VEGF)、肿瘤坏死因子-α (TNF-) 和白细胞介素-1 β (IL-1) 的商业 ELISA 进行了验证，以与用酸性柠檬酸盐-葡萄糖溶液 A (ACD-A) 制备的犬血浆一起使用。用 ACD-A 抗凝的贫血小板血浆 (PPP) 以及用 ACD-A 抗凝并掺入相关犬重组蛋白的 PPP 通过每次 ELISA 进行评估，以计算掺入回收效率。还在另外 2 项测定中评估了加标 PPP 的复制品，以量化测定内和测定间的精度。 TGF-1、PDGF-BB 和 VEGF ELISA 的加标回收效率在预期浓度的 75-125% 范围内。测定内和测定间变异为
Platelet-rich plasma has been studied extensively in dogs, but validation of enzyme-linked immunosorbent assays (ELISAs) for quantifying anabolic growth factors and inflammatory cytokines in canine plasma prepared with citrate-based anticoagulants is not available. We performed a validation of commercial ELISAs for transforming growth factor-beta 1 (TGF-1), platelet-derived growth factor-BB (PDGF-BB), vascular endothelial growth factor (VEGF), tumor necrosis factor-alpha (TNF-), and interleukin-1 beta (IL-1) for use with canine plasma prepared with acid-citrate-dextrose, solution A (ACD-A). Platelet-poor plasma (PPP) anticoagulated with ACD-A as well as PPP anticoagulated with ACD-A and spiked with the relevant canine recombinant proteins were evaluated with each ELISA to calculate the efficiency of spike recovery. Replicates of the spiked PPP were also assessed in 2 additional assays to quantify intra-assay and interassay precision. The efficiency of spike recovery was within 75-125% of the expected concentration for the TGF-1, PDGF-BB, and VEGF ELISAs. The intra- and interassay variability were