Candidates for reference swine serum with anti-Trichinella antibodies
Candidates for reference swine serum with anti-Trichinella antibodies
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DOI：
10.1016/j.vetpar.2015.01.012
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发表时间：
2015-03-15
影响因子：
2.6
通讯作者：
Pozio, Edoardo
中科院分区：
文献类型：
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作者：
Gomez-Morales, Maria Angeles;Ludovisi, Alessandra;Pozio, Edoardo
血清学监测旋毛虫属。在控制系统中饲养的猪的感染被认为是一种可能的诊断工具。然而，没有国际生物标准或参考材料来验证内部检测或商业试剂盒，并提高猪抗旋毛虫IgG血清学检测的实验室间可比性。在这项工作中，潜在的参考血清已经从四个实验感染的猪制备。对血清进行检测、等分、冻干并保持在+4 ℃。由于开发任何参比物质的先决条件之一是计划和执行稳定性研究，因此进行了短期和长期稳定性试验的异构体研究，以评价运输和储存的可能降解效应。任意假设冻干血清样品在+4 ℃下的稳定性。对于短期稳定性研究，将两个装置在20 ℃、+4 ℃、+20 ℃和+50 ℃下储存0、1、2和4周，然后重复检测两次。对于长期稳定性研究，在80 ℃、20 ℃和+4 ℃下储存相同数量的单元和每个单元的重复样品0、6、12、18和24个月。在这两项研究中，随机选择单元样品，并在同一天在重复性条件下进行检测。分析各研究温度下各血清的线性回归与时间，然后检验斜率的显著性。此外，分别计算了1周和3年有效期内短期和长期稳定性的不确定度。对于除一种血清外的所有血清以及除+50 ℃外的所有研究温度，短期稳定性研究的数据表明不存在提示降解的显著趋势。回归线的斜率与零无显著差异。即使血清样品中短期稳定性的不确定度不同，降解速率也被认为是可接受的。对于长期稳定性，两份血清样品的回归线斜率从零开始显著变化，表明储存期间可能存在降解趋势。从长期研究的不确定性中扣除的降解百分比各不相同;然而，两份血清样品在所有测定温度下的降解速率均较低。运送血清样本的最合适温度为20摄氏度、+4摄氏度和+20摄氏度;而血清储存的合适温度为20摄氏度和80摄氏度。(C)2015 Elsevier B. V.版权所有。
Serology to monitor Trichinella spp. infection in pigs reared in controlled system has been claimed as a possible diagnostic tool. However, no international biological standards or reference materials exist to validate in house tests or commercial kits, and to improve the inter-laboratory comparability for the serological detection of anti-Trichinella IgG in pigs. In this work, potential reference sera have been prepared from four experimentally infected pigs. Sera were tested, aliquot, lyophilized, and maintained at +4 degrees C. Since one of the prerequisites for the development of any reference material is to plan and execute stability studies, isochronous studies for short and long term stability testing were carried out to evaluate the possible degradation effects of transportation and storage. The stability of the lyophilized serum samples at +4 degrees C, was arbitrarily assumed. For the short term stability study, two units were stored at 20 degrees C, +4 degrees C, +20 degrees C, and +50 degrees C for 0, 1, 2, and 4 weeks, and then tested in duplicate. For the long term stability study, the same number of units and replicates per unit were stored at 80 degrees C, 20 degrees C, and +4 degrees C for 0, 6, 12, 18 and 24 months. In both studies, unit samples were selected randomly and tested on the same day under repeatability conditions. The linear regression versus time for each serum at each studied temperature was analyzed and then slopes were tested for significance. Further, uncertainty of the short and long term stability was calculated for a shelf life period of one week and three years, respectively. For all sera but one, and for all the studied temperatures but +50 degrees C, the data from the short term stability study indicate the absence of a significant trend that would hint at degradation. The slopes of the regression lines did not significantly vary from zero. Even if the uncertainty of the short term stability was variable among serum samples, the rate of degradation was considered acceptable. For the long term stability, slopes of the regression lines of two serum samples significantly varied from zero, indicating a trend of possible degradation during storage. The percentage of degradation deducted from the uncertainty of the long term study varied; however, two serum samples showed the lower rate of degradation at all the assayed temperatures. The most suitable temperatures for dispatching serum samples are 20 degrees C, +4 degrees C and +20 degrees C; whereas, 20 degrees C and 80 degrees C are suitable temperatures for serum storage. (C) 2015 Elsevier B.V. All rights reserved.