目的对2006年分离自广州登革热患者血清的4株登革1型病毒(GZ57/06、GZ63/06、GZ69/06和GZ128/06)进行全基因组序列测定,并同其他毒株序列进行比对分析,以了解其基因组序列特征及可能的传播来源。方法将登革1型病毒分离株基因组分为9个片段进行RT-PCR扩增,5’与3’末端序列采用RACE法扩增,扩增产物纯化后进行PCR测序。通过末端重叠序列拼接成全长病毒基因组序列,利用生物信息学软件对序列进行比对分析。结果测序获得的4株广州登革1型病毒基因组全长均为10735核苷酸(nt),编码3393个氨基酸。5'和3’端各有一段非编码区,长度分别为94nt和462nt。4株病毒的编码区与非编码区核苷酸及氨基酸序列存在一些差异,其中GZ57/06、GZ63/06和GZ69/06毒株的基因组核苷酸和氨基酸序列同源性均在99%以上,而GZ128/06株变异较其他3株大,编码区差异尤其明显。序列比对分析表明,GZ57/06、GZ63/06和GZ69/06毒株均与泰国2001年分离毒株ThD1/0049/01和ThD1/O102/01同源性最高,而GZ128/06毒株与缅甸1998年分离毒株D1.Myanmar.31987/98同源性最高。这些新分离毒株与东南亚毒株属于同一基因型。结论新分离毒株可能来自泰国等东南亚国家和地区,不同毒株表型差异可能与其基因组特征具有密切关系。
Objective To determine the genome sequences of 4 isolated strains of dengue virus type 1 from dengue patients in Guan gzhou in 2006, and compare them with other strains of dengue virus to identify their genornic characteristics and possible origins. Methods The 9 major genomic fragments of isolated dengue virus type 1 were amplified by RT PCR, and RACE method was employed to amplify the 5' and 3' terminal sequences, the amplified products were purified and then the complete genome sequences were obtained by PCR se- quencing method. The bioinformatic programs were used to analyze the sequences. Results The complete genome of the newly isolated dengue virus type 1 contained 10735 nucleotides in length and coded 3393 amino acids. There were 94 and 462 nucleotides in 5' and 3' non coding region, respectively, and there also were some differences in coding and noncoding regions among these strains. The homology of those strains was over 99% on the sequences of the genome and amino acids among GZA7/06, GZ63/06 and GZ69/06 strain. However, the variation of GZ128/06 strain was more striking than that of other three strains, especially on the coding region of this strain. Sequences a lignment demonstrated that GZ57/06, GZ63/06 and GZ69/06 strains showed the maximal homology with ThD1/0049/01 strain and ThD1/ 0102/01 strain isolated in Thailand in 2001, and GZ128/06 strain showed the maximal homology with D1. Myanmar. 31987/98 strain isola ted in Myanrnar in 1998. The Guangzhou isolates belonged to the same genotype as the strains isolated from Southeast Asia countries. Conclusion The newly isolated dengue virus may originate from Southeast Asian countries and the phenotypic differences of those strains may be related with their genomic characteristics.