云南省中缅边境2015年一起登革热暴发的分子特征分析

郭晓芳; 杨明东; 姜进勇; 李华昌; 朱崇革; 桂琴; 卜力群; 周红宁

Abstract

郭晓芳,杨明东,姜进勇,李华昌,朱崇革,桂琴,卜力群,周红宁.云南省中缅边境2015年一起登革热暴发的分子特征分析[J].Chinese journal of Epidemiology,2016,37(3):398-401

云南省中缅边境2015年一起登革热暴发的分子特征分析

Molecular characteristics of dengue virus outbreak in China-Myanmar border region, Yunnan province, 2015

Received:August 25, 2015

DOI：10.3760/cma.j.issn.0254-6450.2016.03.022

KeyWord: 登革热登革病毒暴发

English Key Word: Dengue fever Dengue virus Outbreak

FundProject:国家自然科学基金(30960327);中缅老越边境地区疟疾/登革热联防联控项目

Author Name	Affiliation	E-mail
Guo Xiaofang	Yunnan Provincial Key Laboratory of Vector-borne Diseases Control and Research and Yunnan Provincial Center of Arborvirus Research of Yunnan Institute of Parasitic Diseases, Pu'er 665000, China
Yang Mingdong	Yunnan Provincial Key Laboratory of Vector-borne Diseases Control and Research and Yunnan Provincial Center of Arborvirus Research of Yunnan Institute of Parasitic Diseases, Pu'er 665000, China
Jiang Jinyong	Yunnan Provincial Key Laboratory of Vector-borne Diseases Control and Research and Yunnan Provincial Center of Arborvirus Research of Yunnan Institute of Parasitic Diseases, Pu'er 665000, China
Li Huachang	Lincang County Center for Disease Control and Prevention, Lincang 677000, China
Zhu Chongge	Gengma County Center for Disease Control and Prevention, Gengma 677500, China
Gui Qin	Gengma County Center for Disease Control and Prevention, Gengma 677500, China
Bu Liqun	Gengma County Center for Disease Control and Prevention, Gengma 677500, China
Zhou Hongning	Yunnan Provincial Key Laboratory of Vector-borne Diseases Control and Research and Yunnan Provincial Center of Arborvirus Research of Yunnan Institute of Parasitic Diseases, Pu'er 665000, China	zhouhn66@163.com

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Abstract:

目的对2015年云南省中缅边境一起登革热暴发查明病因,对流行的登革病毒(DENV)进行分子特征分析,为疾病防控提供病原学证据。方法采用半巢式RT-PCR方法检测2015年7月云南省耿马县孟定镇DENV NS1抗原阳性血清中的DENV特异核酸(CprM基因),对阳性标本用DENV E基因特异引物进行扩增,并将PCR产物送测序,经拼接剪切后的序列在NCBI网站进行BLAST比对,在Megalign中计算核苷酸相似性;在GenBank中选出不同国家和地区不同年度的DENV E基因序列作为参考序列,在Mega 5.05 软件中采用邻接(NJ)法构建系统进化树。结果对25份中国云南省耿马县孟定镇本地病例和14份缅甸输入病例DENV NS1抗原阳性的血清标本进行DENV特异核酸检测,共有21份本地和10份缅甸输入病例标本为DENV-1阳性,其余8份为DENV阴性。测序得到13株(8株来源于本地病例,5株来源于输入病例)1 485 bp的E基因序列,核苷酸相似性为100%,12株(9株来源于本地病例和3株来源于输入病例)406 bp的CprM基因序列,核苷酸相似性为100%。系统进化分析显示,13株E基因序列均属于DENV-1的基因Ⅰ型,位于独立的进化分支。结论本次登革热暴发由DENV-1的基因Ⅰ型引起,该病毒与相邻缅甸区域流行的DENV进化关系最近,当地需加强对登革热防控的综合措施,以防止登革热疫情的进一步扩散。

English Abstract:

Objective To understand the molecular characteristics of a dengue virus outbreak in China-Myanmar border region, Yunnan province, 2015 and provide etiological evidence for the disease control and prevention. Methods Semi-nested RT-PCR was conducted to detect the capsid pre-membrane (CprM) gene of RNA of dengue virus by using dengue virus NS1 positive serum samples collected in Mengdin township, Gengma county, Yunnan province in July, 2015. Some positive samples were then detected by using PCR with specific primers to amplify the full E gene. The positive PCR products were directly sequenced. Then sequences generated in this study were BLAST in NCBI website and aligned in Megalign in DNAstar program. Multiple sequence alignments were carried out by using Mega 5.05 software based on the sequences generated in this study and sequences downloaded from GenBank, including the representative strains from different countries and regions. Phylogenetic trees were constructed by using Neighbor-Joining tree methods with Mega 5.05 software. Results Twenty one of 25 local cases and 10 of 14 imported cases from Myanmar were positive for DENV-1. Eight serum samples were negative for dengue virus. A total of 13 strains with E gene (1 485 bp), including 8 local strains and 5 imported strains, were sequenced, which shared 100% nucleotide sequence identities. Twelve strains with CprM gene (406 bp) from 9 local cases and 3 imported cases shared 100% nucleotide sequence identities. Phylogenetic analyses based on E gene showed that the new 13 strains clustered in genotype Ⅰ of dengue virus and formed a distinct lineage. Conclusions This outbreak was caused by genotypeⅠof DENV-1, which had the closest phylogenetic relationships with dengue virus from neighboring Burma area. Comprehensive measures of prevention and control of dengue fever should be strengthened to prevent the spread of dengue virus.

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