Sequence the complete sequence of bocavirus I with SISPA-PCR.
- 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
- Publication Type:Journal Article
- MeSH: Base Sequence; Bocavirus; genetics; isolation & purification; DNA Primers; genetics; Diarrhea; virology; Genome, Viral; Humans; Parvoviridae Infections; virology; Polymerase Chain Reaction
- From: Chinese Journal of Experimental and Clinical Virology 2010;24(1):14-16
- CountryChina
- Language:Chinese
-
Abstract:
OBJECTIVETo sequence the complete sequence of bocavirus I with sequence independent single primer amplification (SISPA-PCR).
METHODSTo 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 bocavirus was Amplificated with SISPA-PCR, then purificated, cloned and sequenced. The sequences were alighmented in nr with blastn and assembled with DNAstar.
RESULTSA 4834bp sequence of bocavirus I were assembled.
CONCLUSIONSISPA-PCR is an economical and efficient technique for sequence a virus complete genome.
