In an investigation of blood-sucking insects and arboviruses, a virus(YN12243) was isolated from Culicoides samples collected in the Sino-Burmese border region of Yunnan Province, China. The virus caused cytopathic ef...In an investigation of blood-sucking insects and arboviruses, a virus(YN12243) was isolated from Culicoides samples collected in the Sino-Burmese border region of Yunnan Province, China. The virus caused cytopathic effect(CPE) in C6/36 cells and passaged stably. Polyacrylamide gel analysis showed that the genome of YN12243 was composed of 12 segments of double-stranded RNA(dsRNA), with a distribution pattern of 6-6. The nucleotide and amino acid sequences of the coding region(1.12 segments) were17,803 bp and 5,925 amino acids in length, respectively. The phylogenetic analysis of VP1 protein(RdRp) revealed that YN12243 belonged to genus Seadornavirus of family Reoviridae, and further analysis indicated that YN12243 belongs to the Banna virus(BAV) genotype A2. Additionally, YN12243 was located in the same evolutionary cluster as BAV strains isolated from different mosquito species, suggesting that the BAV isolated from Culicoides does not have species barriers. These results indicate that Culicoides can also be a vector for BAV. In view of the hematophagous habits of Culicoides on cattle, horses, deer, and other large animals, as well as the possibility of spreading and causing a variety of animal arboviral diseases, it is important to improve infection detection and monitor the BAV in large livestock.展开更多
目的设计版纳病毒型别特异性测序引物,为构建快速高效经济的版纳病毒全基因组测序平台奠定基础。方法根据Gen Bank已知的版纳病毒基因序列信息进行基因型分析,采用Primer Premier v6.0及Oligo 7.56软件完成版纳病毒亚型特异性扩增引物...目的设计版纳病毒型别特异性测序引物,为构建快速高效经济的版纳病毒全基因组测序平台奠定基础。方法根据Gen Bank已知的版纳病毒基因序列信息进行基因型分析,采用Primer Premier v6.0及Oligo 7.56软件完成版纳病毒亚型特异性扩增引物的设计与评估。使用中国分离的30株版纳病毒进行版纳病毒型别特异性引物工作效率验证和评估。结果通过对30株隶属于2个不同基因亚型的版纳病毒开展全基因组序列测定,获得2套型别工作效率高、扩增效果稳定的基因A型特异性版纳病毒全基因组序列扩增测序引物。基因A1亚型引物一套共26对;基因A2亚型引物一套共计30对。完成了30株版纳病毒全基因组序列扩增。结论设计的版纳病毒型别特异性扩增测序引物可以高效准确地完成版纳病毒全基因组序列扩增,为进一步深入研究版纳病毒分子生物学特征奠定了基础。展开更多
基金supported by the National Natural Science Foundation of China(81290342,81501757)the Special National Project on Research and Development of Key Biosafety Technologies(2016YFC1201904)+1 种基金the National Key Plan for Scientific Research and Development of China(2016YFD0500300)the development grants of State Key Laboratory of Infectious Disease Prevention and Control(2014SKLID103,2015SKLID505)
文摘In an investigation of blood-sucking insects and arboviruses, a virus(YN12243) was isolated from Culicoides samples collected in the Sino-Burmese border region of Yunnan Province, China. The virus caused cytopathic effect(CPE) in C6/36 cells and passaged stably. Polyacrylamide gel analysis showed that the genome of YN12243 was composed of 12 segments of double-stranded RNA(dsRNA), with a distribution pattern of 6-6. The nucleotide and amino acid sequences of the coding region(1.12 segments) were17,803 bp and 5,925 amino acids in length, respectively. The phylogenetic analysis of VP1 protein(RdRp) revealed that YN12243 belonged to genus Seadornavirus of family Reoviridae, and further analysis indicated that YN12243 belongs to the Banna virus(BAV) genotype A2. Additionally, YN12243 was located in the same evolutionary cluster as BAV strains isolated from different mosquito species, suggesting that the BAV isolated from Culicoides does not have species barriers. These results indicate that Culicoides can also be a vector for BAV. In view of the hematophagous habits of Culicoides on cattle, horses, deer, and other large animals, as well as the possibility of spreading and causing a variety of animal arboviral diseases, it is important to improve infection detection and monitor the BAV in large livestock.
文摘目的设计版纳病毒型别特异性测序引物,为构建快速高效经济的版纳病毒全基因组测序平台奠定基础。方法根据Gen Bank已知的版纳病毒基因序列信息进行基因型分析,采用Primer Premier v6.0及Oligo 7.56软件完成版纳病毒亚型特异性扩增引物的设计与评估。使用中国分离的30株版纳病毒进行版纳病毒型别特异性引物工作效率验证和评估。结果通过对30株隶属于2个不同基因亚型的版纳病毒开展全基因组序列测定,获得2套型别工作效率高、扩增效果稳定的基因A型特异性版纳病毒全基因组序列扩增测序引物。基因A1亚型引物一套共26对;基因A2亚型引物一套共计30对。完成了30株版纳病毒全基因组序列扩增。结论设计的版纳病毒型别特异性扩增测序引物可以高效准确地完成版纳病毒全基因组序列扩增,为进一步深入研究版纳病毒分子生物学特征奠定了基础。
