人博卡病毒1型的近似全基因组扩增及HBoV1-4重组分析

Amplification of nearly-complete sequence HBoV1 and recombination analysis among HBoV1-4

目的 扩增人博卡病毒1型的近似全基因组序列,分析HBoV1-4之间的基因重组关系.方法 以检出HBoV1单阳性的鼻咽抽吸物为原始标本,采用PCR的方法扩增HBoV1的5个片段并测序,Blast比对确定是HBoV1片段后用DNAMAN进行序列拼接,拼接序列与已提交的近似全基因组的HBoV1在不同编码区进行同源性分析,并构建进化树；对HBoV1-4的不同编码区进行序列比对和进化分析,揭示HBoV1-4间的基因重组.结果 得到近似全长为5287 bp的HBoV1-NC序列,同源性分析发现HBoV1-NC与重庆株进化关系最近,与广州株进化距离最远；HBoV3可能由HBoV1和HBoV4重组进化而来,HBoV4可能由HBoV2与HBoV3重组进化而来.结论 本研究成功获得近似全基因组的HBoV-NC,HBoV1-4间存在着基因重组关系.

Objective To amplify the nearly-complete sequence of human Bocavirus 1 （HBoV1）and analyze the recombination relationship among HBoV1-4.Methods Five fragments of HBoV1 genome were amplified by PCR from the HBoV1 single positive nasopharyngeal aspirates,then the five fragments were sequenced and analyzed through the blast in nucleotide acid database.The sequence assembly was conducted by DNAMAN,the homogeneous analysis was performed among the HBoV1 sequences that have been published in nucleotide acid database and then created the phylogenetic tree.At last,the recombination relationship was analyzed among HBoV1-4.Results A total 5287 bp length sequence was amplified in this study,which was the nearly-complete sequence of HBoV1 that was named HBoV1-NC.HBoV1-NC had the closest relationship with Chongqing strain,however,had the farthest relationship with Guangzhou strain through the homogeneous analysis.HBoV3 may be a recombinant derived from HBoV1 and HBoV4,HBoV4 may be a recombinant derived from HBoV2 and HBoV3 through recombination analysis of HBoV1-4.Conclusion The nearly-complete sequence HBoV1-NC was amplified in this study and HBoV1-4 had the recombination relationship.