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RT-qPCR assay for detection of mink astrovirus in outbreaks of diarrhea on Danish mink farms

机译:RT-QPCR测定检测丹麦水貂农场爆发的貂皮中毒剂

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摘要

Diarrhea in mink kits is a major cause of disease and mortality in the mink production. The etiology remains unknown in most outbreaks due to a lack of diagnostic assays. In the current study we present an RT-qPCR method to detect mink astrovirus in fecal samples from mink kits with diarrhea. All sampled animals were classified based on age and patoanatomical evaluation as having pre-weaning diarrhea, diarrhea in the growth period or as having no macroscopic signs of diarrhea. Fecal samples were analyzed for MiAstV with RT-qPCR, next generation sequencing and electron microscopy in parallel. Mink astrovirus was detected with RT-qPCR in 92 out of 203 samples. This detection was confirmed by next generation sequencing in a high proportion of samples (22/27), and by visualization of astrovirus particles with EM in some of the samples. Mink astrovirus was highly prevalent (68%) among kits in the outbreaks of pre-weaning diarrhea, in particular outbreaks from May, while less prevalent in outbreaks in June. Mink astrovirus was detected in outbreaks of diarrhea in the growth period, though in a much lesser extent than in the pre-weaning period. The role of mink astrovirus in the diarrhea disease complex of mink remain to be investigated, and for that purpose this sensitive and robust RT-qPCR can be a valuable tool in the future.
机译:貂皮试剂盒的腹泻是貂皮生产中疾病和死亡率的主要原因。由于缺乏诊断测定,病因在大多数爆发中仍然未知。在目前的研究中,我们提出了一种RT-QPCR方法,以检测来自Mink试剂盒的粪便样品中的貂鱼雾病毒。所有取样的动物都是根据年龄和对细胞分析评估进行分类,因为在生长期腹泻,腹泻腹泻或没有腹泻的宏观症状。用RT-QPCR分析MIASTV的粪便样品,下一代测序和电子显微镜并联。用RT-QPCR检测Mink Astovirus,在203个样品中的92中检测到92。通过高比例的样品(22/27),并通过在一些样品中与EM的Astrovirus颗粒的可视化来确认该检测。貂皮里病毒在爆发前腹泻的爆发中具有普遍存在的(68%),特别是从5月份的爆发,而6月份爆发的疫情较少。 Mink Astovirus在生长期爆发中被检测到爆发,但在比后预期期间的程度越来越大。 Mink Astovirus在貂皮中的腹泻疾病复合物中的作用仍有待研究,并且对于此目的,这种敏感和稳健的RT-QPCR将来可以是一个有价值的工具。

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