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A novel RNA-based in situ hybridization to detect Seneca Valley virus in neonatal piglets and sows affected with vesicular disease

机译:一种新颖的基于RNA的原位杂交技术可检测出受水疱病影响的新生仔猪和母猪中的塞内卡谷病毒

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摘要

Seneca Valley virus (SVV) is the causative agent of an emerging vesicular disease in swine, which is clinically indistinguishable from other vesicular diseases such as foot-and-mouth disease. In addition, SVV has been associated with neonatal mortality in piglets. While a commercial SVV qRT-PCR is available, commercial antibodies are lacking to diagnose SVV infections by immunohistochemistry (IHC). Thus, a novel in situ hybridization technique—RNAscope (ISH) was developed to detect SVVRNA in infected tissues. From a total of 78 samples evaluated, 30 were positive by qRT-PCR and ISH-RNA, including vesicular lesions of affected sows, ulcerative lesions in the tongue of piglets and various other tissues with no evidence of histological lesions. Nineteen samples were negative for SVV by qRT-PCR and ISH-RNA. The Ct values of the qRT-PCR from ISH-RNA positive tissues varied from 12.0 to 32.6 (5.12 x 106 to 5.31 RNA copies/g, respectively). The ISH-RNA technique is an important tool in diagnosing and investigating the pathogenesis of SVV and other emerging pathogens.
机译:塞内卡河谷病毒(SVV)是猪中新兴水疱病的病原体,在临床上与其他水疱病(如口蹄疫)没有区别。此外,SVV与仔猪的新生儿死亡率有关。尽管可以使用商业SVV qRT-PCR,但缺乏通过免疫组织化学(IHC)诊断SVV感染的商业抗体。因此,开发了一种新颖的原位杂交技术-RNAscope(ISH)来检测感染组织中的SVVRNA。在总共评估的78个样本中,有30个通过qRT-PCR和ISH-RNA呈阳性,包括受影响母猪的囊泡病变,仔猪舌头的溃疡性病变和其他各种组织,没有组织学病变的迹象。通过qRT-PCR和ISH-RNA对19份SVV阴性。 ISH-RNA阳性组织的qRT-PCR的Ct值从12.0到32.6(分别为5.12 x 10 6 到5.31 RNA拷贝/ g)。 ISH-RNA技术是诊断和调查SVV和其他新兴病原体发病机制的重要工具。

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