摘要
为了解云南省西双版纳傣族自治州(西双版纳州)登革热流行季节城区的媒介伊蚊携带虫媒病毒的种类,对2018年8—10月在西双版纳州城区采集的媒介伊蚊(白纹伊蚊和埃及伊蚊)和7—11月采集的登革病毒(dengue virus,DENV)NS1阳性血清采用实时荧光RT-PCR进行DENV分型检测,用RT-PCR扩增DENV E基因。用C6/36白纹伊蚊细胞对蚊悬液进行病毒分离,对有细胞病变效应(cytopathic effects,CPE)的分离物进行虫媒病毒和昆虫特异病毒RT-PCR检测。对阳性的RT-PCR扩增产物测序后采用BLAST在线将序列与GenBank数据库中相似序列进行比对,用MEGA7进行进化分析。结果从51组伊蚊(共1431只)中检测到2组埃及伊蚊为DENV-1阳性,从54份血清中检测到52份为DENV-1阳性,得到1条来源于埃及伊蚊和27条血清来源的DENV-1 E基因序列,蚊虫来源的E基因与血清来源的E基因核苷酸相似性为97.4%~100.0%,均属于基因Ⅰ型,与近年东南亚国家流行的DENV-1进化关系最近。从分离到的18株有CPE的培养液中鉴定出17株拉蒂纳病毒(La Tina virus,LTNV,属于黄病毒科)和4株砂拉越病毒(Sarawak virus,SWKV,属于T4病毒科),其中3株为LTNV和SWKV同时检测阳性。本研究结果表明,西双版纳州媒介伊蚊中携带DENV、LTNV和SWKV,有必要持续开展虫媒病毒和昆虫特异性病毒的调查和研究。
To understand the arbovirus carried by the vector Aedes mosquito in the urban areas of Xishuangbanna Dai Autonomous Prefecture(Xishuangbanna Prefecture)during the dengue fever epidemic season in Yunnan province,real-time RT-PCR analysis for DENV serotyping and E gene detection were conducted on the RNA extracted from Aedes mosquitoes(including Ae.albopictus and Ae.aegypti)collected from August to October and DENV NS1 positive serum collected from July to November 2018 in Xishuangbanna Prefecture.A phylogenetic analysis was performed on the sequences obtained from the positive amplification products targeting Flaviviridae sp..C6/36 Ae.albopictus cells were used to isolate viruses from mosquito homogenates.Isolates with cytopathic effects(CPE)were identified by arbovirus and insect-specific virus RT-PCR and sequencing.The sequences were compared to the online GenBank database by BLAST.The DENV serotyping detection results indicated that two groups of Ae.aegypti among 51 groups of Aedes mosquitoes(1431 mosquitoes in total)and 52 out of 54 serum samples were positive for DENV-1.One mosquito DENV-1 E gene sequence and 27 serum DENV-1 E gene sequences were obtained.The nucleotide sequence identities of E genes derived between mosquito and serum were 97.4%~100.0%and they belonged to genotype I,which had the closest genetic relationship with DENV-1 from Southeast Asian nations.Additionally,Seventeen La Tina virus(LTNV,belonging to Flaviviridae)strains and four Sarawak virus(SWKV,belonging to Alphateraviridae)strains were identified from 18 isolates using regular CPE isolated from suspensions derived from 51 groups containing vector Aedes mosquitoes,of which three strains were simultaneously positive for both LTNV and SWKV detection.The results from this study indicate that vector Aedes mosquitoes in Xishuangbanna Prefecture carry DENV,LTNV,and SWKV and that further researches on arboviruses and insect-specific viruses is urgent necessary.
作者
郭晓芳
周克梅
陈子龙
许翔
兰学梅
王雪菲
董利民
高阳
李鸿斌
李卫平
GUO Xiao-fang;ZHOU Ke-mei;CHEN Zi-long;XU Xiang;LAN Xue-mei;WANG Xue-fei;DONG Li-min;GAO Yang;LI Hong-bin;LI Wei-ping(Yunnan Institute for Parasitic Disease Control,Yunnan Provincial Key Laboratory of Vector-borne Diseases Control and Research,Puer 665000,Yunnan,China;Xishuangbanna Dai Autonomous Prefecture Center for Disease Control and Prevention,Jinghong 666100,Yunnan,China)
出处
《寄生虫与医学昆虫学报》
CAS
2023年第4期202-209,共8页
Acta Parasitologica et Medica Entomologica Sinica
基金
国家自然科学基金项目(No.81560548)。
