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用于抗体检测的猪血清样本中经典猪瘟病毒的灭活

Inactivation of Classical Swine Fever Virus in Porcine Serum Samples Intended for Antibody Detection.

作者信息

Meyer Denise, Petrov Anja, Becher Paul

机构信息

EU and OIE Reference Laboratory for Classical Swine Fever, Institute of Virology, Department of Infectious Diseases, University of Veterinary Medicine Hannover, Foundation, 30559 Hannover, Germany.

出版信息

Pathogens. 2019 Dec 5;8(4):286. doi: 10.3390/pathogens8040286.

Abstract

Shipping of serum samples that were taken from pigs infected with classical swine fever (CSF) virus is frequently requested with the objective of serological analyses, not only for diagnostic purposes but also for exchange of reference materials that are used as control material of diagnostic assays. On the basis of the fact that an outbreak with CSF is associated with enormous economic losses, biological safety during the exchange of reference material is of great importance. The present study aimed to establish a pragmatic approach for reliable CSF virus (CSFV) inactivation in serum without impairing antibody detection. Considering the fact that complement inactivation through heating is routinely applied, the basic idea was to combine heat treatment with the dilution of serum in a detergent containing buffer in order to facilitate the inactivation process. The results show that treatment of serum samples with phosphate buffered saline-Tween (final concentration = 0.15%) along with incubation at 56 °C for 30 min inactivated CSFV and such treatment with ≤ 0.25% PBS-Tween does not impair subsequent antibody detection by ELISA or virus neutralization test. This minimizes the risk of virus contamination and represents a valuable contribution to a safer CSF diagnosis on a national and international level.

摘要

从感染经典猪瘟(CSF)病毒的猪身上采集的血清样本经常被要求用于血清学分析,不仅用于诊断目的,还用于作为诊断检测对照材料的参考物质的交换。鉴于CSF疫情会造成巨大经济损失,参考物质交换过程中的生物安全至关重要。本研究旨在建立一种实用方法,在不影响抗体检测的情况下可靠地灭活血清中的CSF病毒(CSFV)。考虑到常规采用加热进行补体灭活这一事实,基本思路是将热处理与在含去污剂的缓冲液中稀释血清相结合,以促进灭活过程。结果表明,用磷酸盐缓冲盐水 - 吐温(终浓度 = 0.15%)处理血清样本并在56℃孵育30分钟可灭活CSFV,且使用≤0.25% PBS - 吐温进行此类处理不会损害随后通过ELISA或病毒中和试验进行的抗体检测。这将病毒污染风险降至最低,并为国家和国际层面更安全的CSF诊断做出了宝贵贡献。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/f0a2/6963797/559398f7c022/pathogens-08-00286-g001.jpg

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