青藏高原喜马拉雅旱獭粪便携带腹泻相关病毒的调查分析

Investigation and sequence analysis of diarrhea related viruses in the feces of Marmota himalayana on the Qinghai-Tibet Plateau, China

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 diarrhea associated viruses in the feces of Marmota himalayana on the Qinghai- Tibet Plateau, China, and also analyze these viruses sequences. Methods We designed the primers according the conserved region of genome, and detected 96 fecal samples by direct PCR or one-step RT-PCR; the positive PCR products were sequenced and aligned with the reference strains using PhyML 3.0 software. We also analyzed the evolution of these virus＇ s sequences. Results A total of 32 astroviruses, 42 picobirnaviruses, and 5 kobuviruses were detected in 96 feces of M. himalayana. The positive of these viruses were 35.4%, 43.8%, and 5.2%, respectively. These virus＇s sequences had a nueleotide homology of 78%, 77%, and 93% and the amino acid homology of 77%, 84%, and 91% with the reference strains, respectively. Adenovirus, sapovirus and norovirus were not detected by the degenerate primers. Conclusion There are picobirnaviruses, astroviruses and kobuviruses in the feces of M. himalayana. These studies will provide important clues for further study of viruses in M. himalayana.