摘要
目的对衢州市2017年1例曾有孟加拉国旅行史的发热患者血清标本进行基孔肯雅病毒(CHIKV)全基因组序列测定及遗传进化分析。方法无菌采集患者血液,分离血清后采用实时荧光定量PCR检测CHIKV和登革热病毒核酸,反转录PCR扩增CHIKV全基因组靶向基因片段并测序,采用生物信息学软件(Contig Express、SeqMan、Clustalx V2.0、BioEdit V7.0、GENEDOC和MEGA X)分析全基因序列。结果患者血清为CHIKV核酸阳性。测序数据拼接处理后获得CHIKV全基因组序列(QZ0823株),包含11 787 bp核苷酸,非结构蛋白主要结构位点未发现变异,结构蛋白缺乏E1中的适应性突变A226V和E2中可能影响CHIKV神经毒力的新取代K252Q突变,不存在增强埃及伊蚊的适应性突变E1:K211E和E2:V264A。全基因组序列进化分析显示,QZ0823与孟加拉国、印度、巴基斯坦流行株彼此紧密聚集,具有密切的进化相关性。结论衢州市输入性基孔肯雅病毒QZ0823全基因组和结构基因C-E3-E2-6K-E1核苷酸序列均符合CHIKV特征,与孟加拉国流行株同属东中南非洲遗传谱系(ECSA)。
Objective To perform whole-genome sequencing and phylogenetic analyses of Chikungunya virus(CHIKV)from the serum sample of a febrile patient who had traveled to Bangladesh in Quzhou, 2017. Methods Blood samples were collected aseptically from the patient. The serum was separated for detection of nucleic acids of CHIKV and Dengue virus by RT-PCR. The genome-wide target gene fragments of CHIKV were amplified by RT-PCR and then sequenced. The whole-genome sequence was analyzed by bioinformatic programs: Contig Express, SeqMan, Clustalx V2.0, BioEdit V7.0,GENEDO and MEGA X. Results The patient’s serum was positive for CHIKV nucleic acids. The sequencing data were assembled to obtain the whole-genome sequence of CHIKV strain QZ0823, containing 11 787 bp nucleotides. No mutation was found in major structural sites of non-structural proteins. For structural proteins, E1 lacked the adaptive mutation A226 V;E2 lacked the new substitution mutation K252 Q that might affect the neurovirulence of CHIKV;there were no Aedes aegypti adaptive mutations E1: K211 E or E2: V264 A. The whole genome-based phylogenetic analysis showed that QZ0823 was tightly clustered with the epidemic strains in Bangladesh, India, and Pakistan, suggesting a close evolutionary relationship. Conclusion The nucleotide sequences of whole genome and structural genes C-E3-E2-6 K-E1 of the imported virus in Quzhou show obvious features of CHIKV. QZ0823 and the epidemic strains in Bangladesh are derived from the Eastern, Central, and Southern African genetic lineage.
作者
王晓光
杨瑞军
黄世腾
曹国平
雷永良
叶灵
WANG Xiao-guang;YANG Rui-jun;HUANG Shi-teng;CAO Guo-ping;LEI Yong-liang;YE Ling(Lishui Center for Disease Control and Prevention,Lishui 323000,Zhejiang Province,China;Quzhou Center for Disease Control and Prevention)
出处
《中国媒介生物学及控制杂志》
CAS
2019年第6期621-625,共5页
Chinese Journal of Vector Biology and Control
基金
2019年浙江省医药卫生科技面上项目计划(2019KY237)~~
