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Detection Of Bovine Respiratory Syncytial Virus In Experimentally Infected Balb/c Mice.

机译:在实验感染的Balb / c小鼠中检测牛呼吸道合胞病毒。

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摘要

The present study used an RT-nested-PCR and an immunohistochemistry assay to detect bovine respiratory syncytial virus in tissues from experimentally infected balb/c mice. As a first step, Chicken Embryo Related (CER) cell monolayers infected with the BRSV-25-BR strain isolated in Brazil were used for antigen production. Then, the infected lung and tracheal tissues of female balb/c mice were collected on 3, 5, 7 and 10 days post-infection and submitted to both techniques. Primers specific to F and G genes that amplify fragments of 481 bp and 371 bp, respectively, were used. The BRSV detection was not successful in all of the animals tested. The genomic fragment of the G gene from the organs of some infected mice on all analyzed post-infection days was amplified. However, in the RT-nested-PCR corresponding to the F gene, it was not possible to observe any amplified fragment. This was probably due to the higher sensitivity of the developed technique to amplify the fragment corresponding to the G gene compared to the F gene. Moreover, only three of the lungs collected five days post-infection were positive by immunohistochemistry. To the author's knowledge, this is the first study reporting bovine respiratory syncytial virus detection in balb/c mice after experimental inoculation.
机译:本研究使用RT巢式PCR和免疫组织化学分析法检测实验感染的balb / c小鼠组织中的牛呼吸道合胞病毒。第一步,将在巴西分离到的BRSV-25-BR株感染的鸡胚相关(CER)细胞单层用于抗原生产。然后,在感染后3、5、7和10天收集雌性balb / c小鼠的被感染的肺和气管组织,并接受两种技术。使用对F和G基因具有特异性的引物,分别扩增481 bp和371 bp的片段。在所有测试的动物中,BRSV检测都不成功。在所有分析的感染后天中,从一些被感染小鼠的器官中扩增出G基因的基因组片段。然而,在对应于F基因的RT嵌套PCR中,不可能观察到任何扩增的片段。这可能是由于与F基因相比,已开发技术扩增对应于G基因的片段的敏感性更高。此外,在感染后五天收集的肺中只有三个通过免疫组织化学呈阳性。据作者所知,这是第一项报告在实验接种后在balb / c小鼠中检测牛呼吸道合胞病毒的研究。

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