An immunoturbidimetric assay for bovine haptoglobin.
An immunoturbidimetric assay for bovine haptoglobin.
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DOI：
10.1007/s00580-018-2863-6
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发表时间：
2019-01-01
影响因子：
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通讯作者：
Eckersall, P David
中科院分区：
文献类型：
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作者：
Brady, Nicola;O'Reilly, Emily L;Eckersall, P David
在牛中，血清蛋白结合珠蛋白(HP)是一种主要的急性时相蛋白(APP)，在促炎细胞因子的刺激下，其浓度会上升1000倍以上。因此，这款应用程序是牛感染、炎症和创伤的一个有价值的生物标志物。牛幽门螺杆菌的检测在临床病理和实验研究中变得越来越常见，因为需要天然免疫的生物标记物。最广泛使用的幽门螺杆菌检测方法是利用其与血红蛋白的结合(HP-Hb结合检测)，这种检测方法在低pH值下能够保存血红蛋白中的天然过氧化物酶活性。这项检测适用于所有物种，包括狗、猫和猪等物种，这些物种的健康动物的Hp水平高于健康牛，因此可以自动化的牛特异性免疫分析将是可取的。因此，一种新的自动化物种特异性免疫比浊法(IT)已经被开发出来。验证研究表明，批内和批间变异系数分别低于5%和9%，从添加了牛幽门螺杆菌的样品中回收率为99%，定量限为0.033g/L。该方法不受黄疸或血脂的影响，但受到血红蛋白的适度干扰，与Hp-Hb结合试验显示出显著的相关性。这种针对牛Hp的新型IT检测方法将允许对这一重要的牛APP进行自动分析，以识别当前Hp-Hb结合分析无法检测到的Hp浓度的变化。它将使这种分析能够纳入牛群健康评估、动物福利分析以及用于牛的药物和研究。
In cattle, the serum protein haptoglobin (Hp) is a major acute phase protein (APP) that rises in concentration over a thousand fold following stimulation by pro-inflammatory cytokines. As such, this APP is a valuable biomarker for infection, inflammation and trauma in cattle. The assay for bovine Hp is becoming more commonplace in clinical pathology and in experimental studies when a biomarker of innate immunity is required. The most widely used assay for Hp utilises its binding to haemoglobin (Hp-Hb binding assay), which at low pH enables the preservation of the native peroxidase activity in the haemoglobin. This assay is used for all species, including species such as dog, cat and pig where the level of Hp is higher in healthy animals of these species than in healthy cattle, and therefore a bovine-specific immunoassay that can be automated would be desirable. Thus, a novel-automated species-specific immunoturbidimetric (IT) assay has been developed. Validation studies showed intra- and inter-assay CVs of below 5% and 9% respectively and a recovery of 99% from samples spiked with bovine Hp and a limit of quantification of 0.033g/L. The assay is not affected by icterus or lipaemia but had moderate interference from haemoglobin and showed a significant correlation with the Hp-Hb binding assay. This novel IT assay for bovine Hp will allow automated analysis of this important bovine APP to identify changes in the Hp concentration not detectable by current Hp-Hb binding assays. It will enable the incorporation of this assay into herd health assessments, animal welfare analysis and for bovine medicine and research.