Department of Virology, Wageningen Bioveterinary Research, PO box 65, 8200 AB, Lelystad, The Netherlands; Department of Biochemistry, Centre for Human Metabolomics, North-West University, South Africa.
Department of Virology, Wageningen Bioveterinary Research, PO box 65, 8200 AB, Lelystad, The Netherlands.
J Virol Methods. 2018 Sep;259:50-53. doi: 10.1016/j.jviromet.2018.06.007. Epub 2018 Jun 12.
Since February 1 2011, rinderpest (RP) has been officially declared eradicated worldwide. National authorities have been requested to destroy all their RP related materials. Nonetheless, their national reference laboratories performing real time reverse transcription polymerase chain reaction assays (PCR diagnostics) need RP positive control samples, since some countries still prefer to maintain diagnostic capability for RP for several reasons. In the future, a similar situation will arise for peste des petits ruminants (PPR) as the ambition has been expressed to eradicate PPR. Anticipating on this, we intended to perform qualified PCR diagnostics without use of infectious RPV or PPRV. Therefore, Newcastle disease virus (NDV) with small RNA inserts based on RPV or PPRV sequences were generated and used as positive control material. Recombinant NDVs (recNDVs) were differentially detected by previously established PCR diagnostics for RPV or PPRV. Both recNDVs contain a second PCR target showing that additional targets in NDV are feasible and would increase the diagnostic sensitivity by use of two PCR assays. RecNDV with small PCR targets is not classified as RPV or PPRV containing material, and can be used to mimic RPV or PPRV. Using these recNDVs as virus positive material contributes to the ambition of worldwide eradication, while qualified PCR diagnostics for these OIE-listed diseases remains operational.
自 2011 年 2 月 1 日起，牛瘟（RP）已在全球范围内被正式宣布根除。各国当局被要求销毁其所有与 RP 相关的材料。尽管如此，由于某些国家出于多种原因仍倾向于保持对 RP 的诊断能力，因此其国家参考实验室仍需要进行实时逆转录聚合酶链反应（PCR 诊断）的 RP 阳性对照样本。在未来，小反刍动物疫病（PPR）也将出现类似情况，因为已经表达了根除 PPR 的愿望。有鉴于此，我们打算在不使用传染性 RPV 或 PPRV 的情况下进行合格的 PCR 诊断。因此，我们基于 RPV 或 PPRV 序列生成了带有小 RNA 插入的新城疫病毒（NDV），并将其用作阳性对照材料。通过先前建立的针对 RPV 或 PPRV 的 PCR 诊断方法可以区分出重组 NDV（recNDV）。这两种 recNDV 都包含第二个 PCR 靶标，表明 NDV 中的其他靶标是可行的，并且通过使用两个 PCR 检测可以提高诊断的灵敏度。含有小 PCR 靶标的 recNDV 不被归类为含有 RPV 或 PPRV 的材料，可用于模拟 RPV 或 PPRV。使用这些 recNDV 作为病毒阳性材料有助于实现全球根除的目标，同时也使这些 OIE 列出的疾病的合格 PCR 诊断保持运作。
