Aleutian disease (AD), caused by Aleutian mink disease virus (AMDV), causes significant welfare problems to mink, and financial losses to the farmers. As there is no vaccine or treatment available, reliable diagnostics is important for disease control. Here, we set up a probe-based real-time PCR (NS1-probe-PCR) to detect all strains of AMDV. PCR was validated and compared to two other real-time PCR methods (pan-AMDV- and pan-AMDO-PCR) currently used for AMDV diagnostics in Finland. The NS1-probe-PCR had a similar detection limit of 20 copies/reaction based on plasmid dilution series, and similar or better diagnostic sensitivity, when evaluated using spleen samples from mink, and stool samples from mink and foxes. None of the three PCR tests cross-reacted with other parvoviruses. The NS1-probe-PCR also showed a significantly higher specificity than the pan-AMDO-PCR with spleen samples and the best specificity with stool samples. Furthermore, it produced the results more rapidly than the other two PCRs making it a promising tool for both diagnostic and research purposes.

中文翻译：

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开发和验证诊断阿留申水貂病病毒的核酸测试。

由阿留申水貂病病毒（AMDV）引起的阿留申病（AD），给水貂造成了严重的福利问题，并给农民造成了经济损失。由于没有可用的疫苗或治疗方法，因此可靠的诊断方法对于疾病控制至关重要。在这里，我们建立了基于探针的实时PCR（NS1-probe-PCR），以检测所有AMDV菌株。PCR已得到验证，并与芬兰目前用于AMDV诊断的其他两种实时PCR方法（pan-AMDV-和pan-AMDO-PCR）进行了比较。当使用来自水貂的脾脏样品以及来自水貂和狐狸的粪便样品进行评估时，基于质粒稀释系列，NS1-probe-PCR的检测限相似，为20个拷贝/反应，并且具有相似或更好的诊断敏感性。三种PCR测试均未与其他细小病毒交叉反应。NS1-probe-PCR对脾脏样品的特异性也高于pan-AMDO-PCR，对粪便样品的特异性最好。此外，它产生的结果比其他两个PCR更快，使其成为用于诊断和研究目的的有前途的工具。