贵州少数民族居住集中地域蜱传播病毒的调查

Ticks are small arthropods that live by sucking blood and could transmit more than 80 virus pathogens during the blood sucking process. The obvious characteristic of tick-borne viral diseases is that they are endemic on fixed areas and their distribution depends on the tick vectors. Tick-borne viral diseases usually first circulate among wild animals by ticks and then transmitted to humans through human development activities. Guizhou Province has 3 ethnic group autonomous prefectures, which has unique geographical, climate and human residential environment that fit the abundant presence of ticks and the transmission of tick-borne viral diseases. It is quite possible that there are undiscovered tick-borne viral pathogens over there. In this study, ticks from Guizhou ethnic group autonomous prefectures will be collected. The tick species, distribution area and seasonal change will be analyzed. Real time reverse transcriptase polymerase chain reaction will be used to check the common tick-borne viruses: Crimean-Congo hemorrhagic fever virus, tick-borne encephalitis virus and SFTSV. LC-MS-MS assay, next generation sequencing technique and virus isolation by cell culture and animal inoculation will be used to identify and isolate new tick-borne viruses. We have succeeded to isolate several new viruses from mosquitos and ticks by using similar strategy. The biological characteristics and genotype of the newly identified viruses will be studied and diagnostic reagents will be developed to offer new tool for clinical diagnosis and disease surveillance. Serum samples from humans, house and wild animals who had contacts with ticks will be collected and antibodies checked for corresponding viruses to identify infection. The results obtained from this project will enrich the knowledge on tick-borne viral pathogens and provide new scientific data for the prevention and control of tick-borne viral diseases.

蜱是一种吸血的媒介昆虫, 在吸血过程中可传播多种病毒。蜱传病毒病的发生具有明显的地方性，先在局部野生动物间经蜱循环，随人类开发而传至人类。贵州有三个少数民族自治州，具有独特的地理、气候、人文环境，适合蜱寄生繁殖，极可能存在未知的蜱传病毒。本项目将采集贵州少数民族居住集中地域蜱类，分析鉴定蜱的种类、分布区域及季节变化；使用荧光定量PCR法检测蜱类感染新疆出血热病毒、森林脑炎病毒、发热伴血小板减少综合征病毒等已知蜱传病毒的感染状况；利用LC-MS-MS质谱分析多肽序列结合高通量测序及病毒分离培养方法对未知蜱传病毒进行检测、分离、鉴定（使用类似方案我们已成功从蚊及蜱分离到数个新病毒）；对检测到的病毒进行生物学性状、基因型的系统解析并开发诊断试剂，为临床诊断及疾病监测提供工具；采集接触蜱类的人、家畜及野生动物血清，检测相应病毒抗体以明确感染状况，为蜱传病毒病的预防控制提供理论基础与实验数据。

在贵州省6个市州18个县（市、区）开展了蜱采集工作，在四县设监测点观察蜱分布的季节变化，发现长角血蜱、褐黄血蜱、微小牛蜱是贵州地区主要的蜱种，4月份是蜱高峰季节。实时荧光定量PCR检测未发现蜱传SFTS病毒、蜱传森林脑炎病毒及蜱传新疆出血热病毒阳性蜱。蜱携带细菌培养发现贵州地区蜱体内携带细菌种类繁多，含多种人兽共患病菌；宠物寄生蜱调查发现贵阳宠物蜱体内立克次体阳性率较高，为拉乌尔立克次体和黑龙江立克次体；家畜调查发现黔南州羊感染蜱传无形体率较高；体外表达、纯化了重组蜱传SFTS病毒及同科RFV病毒核衣壳蛋白，制备出了的单克隆抗体，建立了检测SFTS病毒和RFV病毒的血清ELISA和胶体金检测方法；本项目为贵州蜱传播疾病防控提供了理论依据与技术基础，有助于临床医生、疾病防控人员及民众了解蜱传播疾病危害，为蜱传播疾病的诊治与防控提供了技术支撑。

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