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目的对2013年浙江省登革热病例进行实验室确诊与病原的分子溯源。方法对患者血清样本同时进行登革热病毒Ig M抗体和核酸检测及病毒分离,阳性分离株采用RT-PCR方法扩增E基因,进行核苷酸序列测定和同源性与进化分析。结果 18例患者发病前均有在境外登革热流行区暴露史,从患者18份血清样本中检测到登革热病毒Ig M抗体阳性15份(83.3%),核酸阳性8份(44.4%),分离到登革热1型病毒6株,4型1株。6株登革热1型株之间E基因核苷酸和氨基酸同源性分别为90.9%~99.7%和96.8%~100%,与浙江省2004年登革热1型株之间核苷酸和氨基酸同源性分别为91.4%~98.5%和96.6%~99.0%;E基因进化树显示,登革热1型株位于GⅠ、GⅣ和GⅤ基因亚型上,登革热4型株位于GⅠ亚型上,2013年7株登革热病毒与浙江省2004-2009年登革热病毒之间亲缘关系较远。结论证实2013年浙江省登革热病例均为输入性,输入国为菲律宾、安哥拉、斯里兰卡和巴西等;登革热1型株基因亚型分别为亚洲型、南太平洋型和美洲/非洲型,登革热4型株为东南亚型;登革热4型病毒为浙江省首次分离。
Objective To diagnose and trace the origin of dengue in Zhejiang Province in 2013. Methods The dengue virus IgM antibody and nucleic acid were detected and the virus was isolated from the serum of patients. The positive isolates were amplified by RT-PCR and sequenced. The nucleotide sequences were determined and their homology and evolution were analyzed. Results Before the onset of the disease, all the 18 patients were exposed to the dengue epidemic area outside of the country. Fifteen positive (83.3%) and 18 (44.4%) were positive for dengue virus IgM antibodies from 18 serum samples. Dengue fever 6 strains of type 1 virus and 1 strain of type 4. The nucleotide and amino acid homology of E gene between 6 strains of dengue type 1 were 90.9% -99.7% and 96.8% -100%, respectively, which were homologous to nucleotide and amino acid of dengue type 1 strain in Zhejiang Province in 2004 Were 91.4% -98.5% and 96.6% -99.0% respectively. The E gene phylogenetic tree showed that the dengue type 1 strain was located on the GⅠ, GⅣ and GⅤ genotypes, the dengue 4 type strain was on the GⅠ subtype, and that in 2013 was 7 strains Dengue virus and Zhejiang province 2004-2009 dengue virus distantly related. Conclusion The cases of dengue fever in Zhejiang Province in 2013 were all imported. The importing countries were Philippines, Angola, Sri Lanka and Brazil. The genotypes of dengue type 1 strains were Asian, South Pacific and American / Africa, and Dengue 4 For the Southeast Asian type; dengue 4 virus isolated for the first time in Zhejiang Province.