Almeida Renata Servan, Domingues Helena Gallicchio, Coswig Lia Treptow, D'Arce Regina Celia Freitas, de Carvalho Rodrigo Franco, Arns Clarice Weis
Laboratório de Virologia Animal, Departamento de Microbiologia e Imunologia, Instituto de Biologia, Universidade Estadual de Campinas-UNICAMP, 13081-970 Campinas, SP, Brazil.
Vet Res. 2004 Mar-Apr;35(2):189-97. doi: 10.1051/vetres:2004003.
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.
本研究采用逆转录巢式聚合酶链反应（RT-nested-PCR）和免疫组织化学检测法，对实验感染的Balb/c小鼠组织中的牛呼吸道合胞病毒进行检测。第一步，使用感染了巴西分离的BRSV-25-BR毒株的鸡胚相关（CER）细胞单层来生产抗原。然后，在感染后第3、5、7和10天收集雌性Balb/c小鼠的感染肺组织和气管组织，并进行这两种检测技术。使用了分别扩增481 bp和371 bp片段的F基因和G基因特异性引物。并非所有受试动物的BRSV检测都成功。在所有分析的感染后天数，部分感染小鼠器官中G基因的基因组片段得到了扩增。然而，在对应F基因的RT-nested-PCR中，未观察到任何扩增片段。这可能是由于所开发的技术对扩增G基因对应片段的敏感性高于F基因。此外，感染后5天收集的肺组织中，只有3个通过免疫组织化学检测呈阳性。据作者所知，这是首次报道实验接种后在Balb/c小鼠中检测到牛呼吸道合胞病毒的研究。
