深圳市首例本地登革热3型病例的病原学检测与分析  

The etiology detection and analysis for the first local case of dengue virus type 3 infection in Shenzhen

作　　者：阳帆[1] 王敬忠[1] 黄亚兰[1] 吴春利[1] 黄达娜[1] 李玥[1] 唐屹君[1] 张仁利[1] 

机构地区：[1]深圳市疾病预防控制中心微生物检验科,518055

出　　处：《国际病毒学杂志》2017年第4期264-269,共6页International Journal of Virology

基　　金：深圳市科创委科技研发资金基础研究项目（JCYJ20150402102135500）

摘　　要：目的对2016年深圳市报告的首例本地疑似登革热病例查明病因.分离鉴定病原体,从分子水平分析分离株的生物学特征.方法对疑似患者血清标本采用ELISA、胶体金免疫层析法和荧光RT-PCR方法分别检测登革病毒抗体、NS1抗原和病毒核酸,并用C6/36细胞分离登革病毒.采用RT-PCR方法扩增病毒PrM/M-E基因后进行序列测定,并与不同国家和地区的登革毒株进行同源性比较和进化树分析.结果从患者血清标本中检测到登革病毒IgM抗体、NS1抗原和登革3型病毒核酸,并成功分离到登革3型病毒,将其命名为DENV3-SZ1648.深圳市登革3型病毒分离株SZ1648与登革3型国际标准株H87株、国内外流行株80-2、GWL-25株在PrM/M-E基因上核苷酸同源性分别为92.0％、91.8％和90.3％,而与登革1、2、4型国际标准株HAWAII、NGC、H241同源性分别为68.7％、64.2％和63.2％.进化树显示SZ1648株与2007年印度尼西亚分离株MKS-0098亲缘关系最近,在进化树的同一分支上,和85-159株（Indonedia 1985）、2167株（Tahiti 1989）、29472株（Fiji1992）等同属基因Ⅰ亚型.患者发病前1个月在深圳居住,无输血史、无外出史.结论从病原学、血清学和分子生物学特征上均证实该本地病例是由登革3型病毒引起,这也是深圳市首次报道存在本地登革3型病毒的疫情,该毒株最有可能来源于印度尼西亚.Objective To determine the etiology of the first local dengue fever case reported in Shenzhen city in 2016,and to isolate,to identify as well as to study the molecular characteristics of the isolated dengue virus strain.Methods The serum sample collected from the suspect dengue fever case was detected for IgM,IgG by ELISA,and NS1 antigen by immunochromatography and dengue virus nucleic acid by real-time RT-PCR.The samples were further inoculated in C6/36 cells for virus isolation.PrM/M-E gene of isolated virus strain was amplified by RT-PCR and sequenced to construct homology comparison and phylogenetic tree of PrM/M-E gene of Shenzhen dengue virus with the strains isolated from other areas.Results IgM,NS1 antigen and RNA of dengue virus type 3 were detected in the serum sample.Type 3 dengue virus named DENV3-SZ1648 was successfully isolated from the serum sample.The homology of nucleotide sequence of PrM/M-E gene of SZ 1648 with standard type 3 dengue virus H87 strain,80-2 strain,GWL-25 strain were 92.0%,91.8% and 90.3%,respectively.The homology with standard dengue virus 1,2,4 in the same fragment were 68.7%,64.2% and 63.2%,respectively.The phylogenetic tree indicated that SZ 1648 had the greatest similarity with the MKS-0098 strain （Indonedia 2007） and they lied in the same branch.The isolated dengue virus type 3 belonged to genotype I as the 85-159 strain （Indonedia 1985）,2167 strain （Tahiti 1989） and 29472 strain （Fiji 1992）.The patient lived in Shenzhen and had no history of travelling outside the area in one month before infection.Conclusions The virological,serological and molecular features showed that the local case of dengue fever was caused by dengue virus type 3 and was most likely imported from Indonesia.This is also the first report of the local dengue virus type 3 infection case in Shenzhen.

关 键 词：本地病例 登革3型病毒 PrM/M-E基因 序列分析 

分 类 号：R446.6[医药卫生—诊断学] R512.8[医药卫生—临床医学]