论文部分内容阅读
目的对2017年深圳市龙华区一起输入性登革热病原体进行分子遗传学分析。方法利用C6/36细胞从病人血清中分离登革热病毒(Dengue virus,DENV),对分离得到的DENV的E基因进行测定和系统发生树构建,结合现场流行病学资料对其分子遗传学进行分析。结果成功分离得到1株DENV-1,命名为SZLH_17JB00855_2017。E基因的扩增获得两条目的产物,分别为1 010 bp和829 bp。目的产物经测序拼接后,最终得到长为1 485 bp DNA序列;NCBI数据库多序列在线比对结果显示SZLH_17JB00855_2017毒株E基因与越南2007、2008及2011年分离毒株E基因序列的覆盖率在99%~100%之间;系统发生树分析结果显示,SZLH_17JB00855_2017与VNBID-V1686/2007、VNBID-V1691/2007、VNBID-V3908/2008、VNBID-V3909/2008、VN09DX879/2011亲缘性最近,与近年来广东省各地市暴发的分离株亲缘性较远。结论现场流行病资料和实验室分子遗传学分析都提示本案例报告的登革热病例属于输入性病例,输入地极有可能是越南。
Objective To carry out molecular genetic analysis of imported dengue pathogens together in 2017 in Longhua District, Shenzhen City. Methods Dengue virus (DENV) was isolated from the serum of patients with C6 / 36 cells. The E gene of DENV was isolated and the phylogenetic tree was constructed. Based on the field epidemiological data, molecular genetics was analyzed. Results A strain of DENV-1 was successfully isolated and named as SZLH_17JB00855_2017. E gene amplification obtained two target products, respectively 1 010 bp and 829 bp. After sequencing, the final 1 485 bp DNA sequence was obtained. The online sequence comparison of multiple sequences in NCBI database showed that the coverage of the gene sequence of S gene between SZLH_17JB00855_2017 and Vietnam in 2007, 2008 and 2011 was 99 The phylogenetic tree analysis showed that SZLH_17JB00855_2017 has the closest relationship with VNBID-V1686 / 2007, VNBID-V1691 / 2007, VNBID-V3908 / 2008, VNBID-V3909 / 2008 and VN09DX879 / 2011. The outbreaks of isolates from various cities in Guangdong Province are of relatively distant origin. Conclusions Both epidemic data and molecular molecular genetic analysis in the laboratory suggest that the dengue cases reported in this case belong to the imported cases and the imported ones are most likely to be Vietnam.