Sequence the complete sequence of bocavirus I with SISPA-PCR

VernacularTitle:应用不依赖序列的单引物扩增技术获得人博卡病毒全基因序列
Author: Jin-Song LI ¹ ; Wei-Xia CHENG ; Dong-Ping YAO ; Bei LAN ; Jie-Mei YU ; Yan LIU ; Yong-Qing LI ; Qing ZHANG ; Miao JIN ; Zi-Qian XU ; Dan-Di LI ; Zhao-Jun DUAN
Author Information

1. 中国疾病预防控制中心
Keywords: Polymerase chain reaction; Parvoviridae; Sequence analysis
From: Chinese Journal of Experimental and Clinical Virology 2010;24(1):14-16
CountryChina
Language:Chinese
Abstract: Objective To sequence the complete sequence of bocavirus I with sequence independent single primer amplification (SISPA-PCR). Methods To exclude the co-effection samples, all clinical samples of diarrhea cases were screened with special primers of rotavirus, astrovirus, adenovirus, calicivirus and bocavirus I . The virus were enriched through ultracentrifugation. Other nucleic acids,such as human and bacteria genomes, were degradated by DNase I and RNase. DNA of hocavirus was Amplificated with SISPA-PCR,then purificated ,cloned and sequenced. The sequences were alighmented in nr with blastn and assembled with DNAstar. Results A 4834bp sequence of hocavirus I were assembled. Conclusion SISPA-PCR is an economical and Efficient technique for sequence a virus complete genome.