Sequence the complete sequence of bocavirus I with SISPA-PCR / 中华实验和临床病毒学杂志

ABSTRACT

<p><b>OBJECTIVE</b>To sequence the complete sequence of bocavirus I with sequence independent single primer amplification (SISPA-PCR).</p><p><b>METHODS</b>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 bocavirus was Amplificated with SISPA-PCR, then purificated, cloned and sequenced. The sequences were alighmented in nr with blastn and assembled with DNAstar.</p><p><b>RESULTS</b>A 4834bp sequence of bocavirus I were assembled.</p><p><b>CONCLUSION</b>SISPA-PCR is an economical and efficient technique for sequence a virus complete genome.</p>