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目的调查我国青藏高原喜马拉雅旱獭粪便中腹泻相关病毒的携带情况,并对检出病毒进行序列分析。方法根据腹泻相关病毒基因组保守区设计兼并引物,采用直接PCR或一步法RT-PCR方法对96份喜马拉雅旱獭粪便标本核酸进行检测;阳性PCR产物测序后与参考毒株进行序列比对;利用Phy ML 3.0软件对喜马拉雅旱獭粪便中病毒序列进行进化分析。结果 96份青藏高原喜马拉雅旱獭粪便中共检出星状病毒32份、小双节RNA病毒42份和嵴病毒5份,阳性率分别为35.4%、43.8%和5.2%;星状病毒、小双节RNA病毒和嵴病毒与参考毒株核苷酸一致性分别为78%、77%和93%,氨基酸一致性分别为77%、84%和91%。兼并引物未检测到腺病毒、札如病毒和诺如病毒。结论青藏高原喜马拉雅旱獭粪便中存在星状病毒、小双节RNA病毒和嵴病毒,为后续喜马拉雅旱獭携带病毒谱系研究提供了重要线索。
Objective To investigate the carriage of diarrhea-associated virus in the Marmota manure from the Qinghai-Tibet Plateau and analyze the sequence of the detected virus. Methods According to the conserved region of diarrhea-associated virus, degenerated primers were designed and 96 samples of D. himalayan manchurian stool samples were detected by direct PCR or one-step RT-PCR. The positive PCR products were sequenced and compared with the reference strains. Phy ML 3.0 Software Evolutionary analysis of virus sequences in the manure of Marmota marmota. Results A total of 96 astroviruses were detected in 96 manure samples from the Himalayan marmot on the Tibetan Plateau. The positive rates of the two samples were 35.4%, 43.8% and 5.2% respectively. Astrovirus, Nucleotide identities of RNA viruses and ridges and reference strains were 78%, 77% and 93%, respectively, and amino acid identities were 77%, 84% and 91%, respectively. Adenovirus, Sapporo virus and Norovirus were not detected by annexing primers. Conclusion The presence of astroviruses, small double-stranded RNA viruses and ridges viruses in the Hamster larvae faeces on the Qinghai-Tibetan Plateau provides important clues for the follow-up study of the virus-carrying lineages in the Himalayan marmot.