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在一项小规模实验研究中,对血清、FTA滤干血和口腔液作为用于猪繁殖与呼吸综合征病毒(PRRSV)诊断的反转录定量聚合酶链反应(RT-qPCR)样本材料进行比较评估。

Comparative evaluation of serum, FTA filter-dried blood and oral fluid as sample material for PRRSV diagnostics by RT-qPCR in a small-scale experimental study.

作者信息

Steinrigl Adolf, Revilla-Fernández Sandra, Wodak Eveline, Schmoll Friedrich, Sattler Tatjana

出版信息

Berl Munch Tierarztl Wochenschr. 2014 May-Jun;127(5-6):216-21.

Abstract

Recently, research into alternative sample materials, such as oral fluid or filter-dried blood has been intensified, in order to facilitate cost-effective and animal-friendly sampling of individuals or groups of pigs for diagnostic purposes. The objective of this study was to compare the sensitivity of porcine reproductive and respiratory syndrome virus (PRRSV)-RNA detection by reverse transcription quantitative real-time PCR (RT-qPCR) in serum, FTA filter-dried blood and oral fluid sampled from individual pigs. Ten PRRSV negative pigs were injected with an EU-type PRRSV live vaccine. Blood and oral fluid samples were taken from each pig before, and 4, 7, 14 and 21 days after vaccination. All samples were then analyzed by PRRSV RT-qPCR. In serum, eight often pigs tested RT-qPCR positive at different time points post infection. Absolute quantification showed low serum PRRSV-RNA loads in most samples. In comparison to serum, sensitivity of PRRSV-RNA detection was strongly reduced in matched FTA filter-dried blood and in oral fluid from the same pigs. These results indicate that with low PRRSV-RNA loads the diagnostic sensitivity of PRRSV-RNA detection by RT-qPCR achieved with serum is currently unmatched by either FTA filter-dried blood or oral fluid.

摘要

最近,对替代样本材料(如口腔液体或滤干血)的研究不断加强,以便以经济高效且对动物友好的方式对猪个体或猪群进行采样用于诊断目的。本研究的目的是比较逆转录定量实时PCR(RT-qPCR)检测血清、FTA滤干血和从个体猪采集的口腔液体中猪繁殖与呼吸综合征病毒(PRRSV)-RNA的敏感性。10头PRRSV阴性猪被注射了一种欧盟型PRRSV活疫苗。在接种疫苗前以及接种后4、7、14和21天从每头猪采集血液和口腔液体样本。然后所有样本通过PRRSV RT-qPCR进行分析。在血清中,8头猪在感染后的不同时间点检测RT-qPCR呈阳性。绝对定量显示大多数样本中血清PRRSV-RNA载量较低。与血清相比,在匹配的FTA滤干血和来自同一猪的口腔液体中,PRRSV-RNA检测的敏感性大幅降低。这些结果表明,在PRRSV-RNA载量较低时,目前血清RT-qPCR检测PRRSV-RNA的诊断敏感性是FTA滤干血或口腔液体无法比拟的。

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