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牛奶浓缩物通过间接 ELISA 提高蓝舌病抗体检测。

Milk concentration improves Bluetongue antibody detection by use of an indirect ELISA.

机构信息

Institute of Virology and Immunoprophylaxis, Sensemattstrasse 293, 3147 Mittelhaeusern, Switzerland.

出版信息

Vet Microbiol. 2010 Jul 14;143(2-4):179-83. doi: 10.1016/j.vetmic.2009.11.036. Epub 2009 Dec 4.

Abstract

A national Bluetongue antibody surveillance in cattle through bulk milk was conducted in Switzerland between July 2007 and June 2008. Using ID Screen Bluetongue Milk ELISA (ID VET, Montpellier, France), samples from 15 out of 210 dairy farms at least once gave a positive result. In only three of these herds bluetongue positive animals were found. Therefore, specificity for bulk milk was not as good as expected and when individual milk samples were tested, it was even lower. As further investigations of positive results were time-consuming and no other ELISA was available at that time, we aimed at discriminating false from true positive samples with a confirmatory test using a protein precipitation method followed by retesting with the same ELISA. Additionally, we examined whether testing of single milk samples can reliably be used to assess status of cows, and whether sampling at the beginning or at the end of milking, as well as freezing and thawing of the milk could influence the performance of the test. Screening with ID VET milk ELISA and confirmatory testing after protein precipitation yielded a clear increase of specificity without any loss of sensitivity in both bulk and single milk samples. This testing scheme allowed minimizing follow-up investigations by blood testing. Antibody levels in plasma and milk showed a good correlation. Tested by logistic regression, none of the possible influencing factors (time point of sample collection, freezing, or milk content of the samples) had a significant influence on the test performance.

摘要

2007 年 7 月至 2008 年 6 月期间,瑞士对全国牛群的蓝舌病抗体进行了抽样调查,采用的是 ID Screen Bluetongue Milk ELISA(ID VET,蒙彼利埃,法国)。在 210 个奶牛场中,有 15 个奶牛场的至少一次抽样检测结果呈阳性。然而,在这些牛群中,仅有 3 个牛群发现了蓝舌病阳性动物。因此,批量奶的特异性并不如预期的那样好,而当对个体牛奶样本进行测试时,特异性甚至更低。由于对阳性结果的进一步调查既耗时又费力,而且当时没有其他 ELISA 可用,我们的目标是通过使用蛋白质沉淀方法进行确认性测试来区分假阳性和真阳性样本,然后用相同的 ELISA 进行复测。此外,我们还研究了在单一牛奶样本测试中是否可以可靠地评估奶牛的状态,以及在挤奶开始或结束时、以及冷冻和解冻牛奶是否会影响测试的性能。使用 ID VET 牛奶 ELISA 进行筛查,然后在蛋白质沉淀后进行确认性测试,在批量和单一牛奶样本中均显著提高了特异性,而没有任何敏感性损失。这种测试方案可以通过血液测试来最小化后续调查。血浆和牛奶中的抗体水平相关性良好。通过逻辑回归测试,样本采集时间、冷冻或样本牛奶含量等可能的影响因素均未对测试性能产生显著影响。

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