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用于检测丹麦水貂养殖场腹泻暴发中貂源星状病毒的 RT-qPCR 检测方法。

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

机构信息

Department of Veterinary and Animal Sciences, Faculty of Health and Medical Sciences, University of Copenhagen, Frederiksberg C, Denmark.

Department of Microbiology, National Veterinary Institute, Uppsala, Sweden.

出版信息

PLoS One. 2021 May 26;16(5):e0252022. doi: 10.1371/journal.pone.0252022. eCollection 2021.

Abstract

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.

摘要

水貂仔兽腹泻是水貂养殖中的主要疾病和死亡原因。由于缺乏诊断检测方法,大多数暴发的病因仍然未知。在本研究中,我们提出了一种 RT-qPCR 方法,用于检测腹泻水貂仔兽粪便样本中的水貂星状病毒。所有采样动物均根据年龄和病理解剖评估分为早发性腹泻、生长期腹泻或无肉眼腹泻迹象。粪便样本同时进行 RT-qPCR、下一代测序和电子显微镜分析以检测 MiAstV。在 203 个样本中,92 个样本通过 RT-qPCR 检测到水貂星状病毒。在大量样本(22/27)中通过下一代测序确认了这种检测,并且在一些样本中通过 EM 观察到星状病毒颗粒进行了确认。水貂星状病毒在早发性腹泻暴发的仔兽中高度流行(68%),特别是 5 月的暴发,而在 6 月的暴发中则较少见。水貂星状病毒在生长期腹泻暴发中被检测到,但程度远低于早发性腹泻。水貂星状病毒在水貂腹泻疾病中的作用仍有待研究,为此,这种敏感而稳健的 RT-qPCR 将来可能是一种有价值的工具。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/f527/8153481/6dad61b8de1c/pone.0252022.g001.jpg

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