南方五省口岸蚊类携带虫媒病毒调查

[目的]掌握南方口岸蚊媒携带病毒的本底资料,为蚊传疾病的预防控制工作提供依据。[方法]采用电动吸蚊器人工法和捕蚊磁场自动法采集南方5省口岸各类蚊虫。采集到的蚊类超低温送至实验室,研磨处理后用荧光PCR方法检测登革病毒、乙脑病毒、黄热病毒、西尼罗病毒、基孔肯雅病毒等重要蚊媒病毒,结果阳性的标本进一步进行PCR扩增和核苷酸序列测定分析;蚊标本研磨液同时用C6／36细胞进行虫媒病毒分离培养,出现细胞病变后分别用黄病毒科、甲病毒科各自的通用引物进行鉴定;对未能鉴定的未知病毒进一步用随机PCR方法进行扩增、克隆、序列测定、Blast搜索。[结果]从南方5省口岸采集到各类蚊虫12575只,鉴定后共分成254组。各组标本经荧光PCR方法检测,结果登革病毒、黄热病毒、西尼罗病毒、基孔肯雅病毒均为阴性;检测到2份福建省来源三带喙库蚊的标本乙脑病毒核酸阳性,经乙脑病毒E基因引物PCR扩增、测序分析证实为GⅠ型病毒。254份标本经C6／36细胞分离培养出现42份细胞病变,用黄病毒科、甲病毒通用引物PCR扩增,均未得到特异片段。选取1份典型病变的细胞培养物进行随机PCR鉴定,结果发现了1种潜伏于C6／36细胞中的浓核病毒。[结论]南方5省口岸蚊媒中可能未携带登革病毒、黄热病毒、西尼罗病毒、基孔肯雅病毒等重要蚊媒病毒,只有少量蚊虫携带乙脑病毒,蚊虫体内检测到的GⅠ型乙脑病毒属于福建省首次发现,出现病变的C6／36细胞可能是由自身潜伏的1种C6／36细胞浓核病毒引起。     

重组酶介导扩增方法快速检测黄热病毒

目的本研究采用重组酶介导的等温核酸扩增方法(RAA),通过使用逆转录酶,建立黄热病毒的一步法等温核酸扩增(RT-RAA)方法。方法根据黄热病毒基因组保守序列设计引物和探针,建立并分析RT-RAA的重复性、特异性、灵敏度;以所建立方法对黄热病毒样本进行检测,同时以基因测序进行验证。结果黄热病毒RT-RAA扩增,体系中加入40 U的逆转录酶扩增效果最佳。该方法检测时间短(20 min),并且灵敏度高,检测下限可达100 copy,与登革病毒、西尼罗病毒、日本乙型脑炎病毒、基孔肯雅热病毒等蚊媒病毒无交叉反应,具有良好的特异性。结论构建的黄热病毒RT-RAA方法具有快速、特异以及灵敏的特点,适应于黄热病毒的口岸快速检测。     

裂谷热病毒荧光定量PCR检测方法的建立

目的建立一种快速、敏感、特异的实时荧光定量PCR方法,用于裂谷热病毒的检测。方法通过序列对比在裂谷热病毒L基因保守区设计引物及Taqman探针,建立实时荧光定量PCR反应体系。结果经优化的荧光定量PCR方法有较好的灵敏度和特异性,对阳性对照质粒标准品的灵敏度可达37拷贝/μl,通过检测同为蚊媒传播的日本脑炎病毒、黄热病毒、登革病毒、基孔肯亚病毒无交叉反应。结论本方法的建立在国境口岸传染病的防控方面有较好的应用前景。     

登革病毒逆转录重组酶聚合酶扩增-测流层析试纸条检测方法的建立与评价

目的建立快速简便的登革病毒检验方法对控制登革热疫情至关重要。方法本研究首次建立了一种登革病毒的RT-RPA-LFD检测技术,结果本研究建立的RT-RPA-LFD检测技术能同时检测四种血清型的登革病毒,具有较好的重复性,特异性高,对乙型脑炎病毒、基孔肯雅热病毒、黄热病毒和塞卡病毒没有交叉检测,并且具有较高的检测灵敏度,对四种血清型的登革病毒检测限都低至10拷贝/反应。结论综合考虑其快速性和操作简便性,RT-RPA-LFD是一种非常有前景的登革病毒一线检测技术。     

多种病毒集合检测芯片的研制及其在临床血清标本检测中的初步应用

目的 研制黄热病、西尼罗热、登革热、基孔肯雅热、埃博拉出血热、马尔堡出血热、拉沙热等多种病毒性传染病集合检测基因芯片,并建立一种适用于直接检测核酸含量较低临床血清标本的新型芯片靶基因扩增标记方法.方法 设计并筛选出上述病毒70mer寡核苷酸特异探针各20条,打印于同一基因芯片上；以phi29 DNA聚合酶结合带标签序列的随机引物进行临床标本中病毒全基因组扩增,再以Cy3荧光染料标记的标签序列引物进行PCR随机扩增标记,标记产物用多种病毒芯片进行杂交检测.结果 检测1～4型登革热、基孔肯雅热病例临床血清标本,结果显示该方法准确、特异、敏感；检测西尼罗热、黄热病、埃博拉出血热、马尔堡出血热、拉沙热等病毒核酸的模拟血清标本,同样得到特异的阳性杂交信号,与预期结果一致.结论 本研究建立的多种病毒集合检测基因芯片及其靶基因扩增标记方法,可直接应用于血清标本中上述病毒核酸的检测.     

基孔肯亚热、克里米亚-刚果出血热、裂谷热病毒核酸液相芯片检测方法的建立

目的建立基孔肯亚热、克里米亚-刚果出血热、裂谷热病毒核酸液相芯片检测方法。方法建立探针偶联至荧光微球、多重PCR扩增方法、杂交检测方法,并对所建立的液相芯片检测方法的灵敏度和特异性进行评价。结果建立的液相芯片检测方法能对基孔肯亚热、克里米亚-刚果出血热、裂谷热病毒中的任意1种、任意2种或者3种同时进行筛查检测,基孔肯亚热病毒检测的灵敏度为1×104copies/PCR,克里米亚-刚果出血热病毒检测的灵敏度为1×105copies/PCR,裂谷热病毒检测的灵敏度为1×103 copies/PCR。该方法对汉坦病毒、埃博拉病毒、黄热病毒、西尼罗病毒、马尔堡病毒的检测结果均为阴性。结论该方法具有高通量、多重检测、快速、敏感、特异的特点,为基孔肯亚热、克里米亚-刚果出血热、裂谷热病毒的筛查检测提供一种新方法。     

大连口岸首例输入性基孔肯雅热病例的实验室检测和确认

目的对大连口岸1名入境发热者的血液样本进行检测和确认,为输入性基孔肯雅热疫情的处置提供技术支撑。方法利用核酸检测方法对入境旅客血液样本进行基孔肯雅病毒、登革病毒、寨卡病毒、黄热病毒及疟原虫等虫媒传染病病原体进行筛查。结果经检测确认,该样本基孔肯雅病毒核酸阳性,特异性引物扩增获得序列与已知基孔肯雅病毒核酸序列同源性达99.61%。结论大连口岸存在基孔肯雅热输入风险,应加强国境口岸基孔肯雅热防控。     

福州口岸入境船舶上首次发现埃及伊蚊及其携带的病原体

[目的]通过加强入境船舶医学媒介监测,防止新蚊种、蚊媒传染病传入和传播。[方法]对入境船舶的积水容器进行蚊幼虫监测,并采样送实验室进行形态学鉴定,登革热、黄热病、基孔肯雅热等病原检测,对船舶实施灭蚊等卫生处理,对船员施行流行病学调查、体温检测、医学检查等措施。[结果]全船发现9处积水,1处蚊幼虫阳性,积水4000ml,幼虫70条,羽化后经实验室鉴定该蚊幼虫均为埃及伊蚊,未检出登革热、基孔肯雅、西尼罗、乙脑等病毒。对该轮船员进行流行病学调查、医学检查,未发现登革、基孔肯雅热、乙型脑炎、西尼罗热、黄热病等病人。对船舶实施灭蚊等卫生处理措施后,未再发现成蚊和蚊幼虫。[结论]埃及伊蚊属福州地区外来蚊种,是传播黄热病、登革热等热带病的重要媒介。提示福州口岸应提高防范意识,特别应加强来自蚊媒传染病疫区船舶的蚊媒监测、卫生处理工作。     

多种蚊媒传染病病原体悬液芯片检测及分型方法的建立

目的建立同时检测登革病毒(DENV1~4型)、日本脑炎病毒(JEV)、西尼罗病毒(WNV)、黄热病毒(YFV)、基孔肯雅病毒(CHIKV)、间日疟原虫(Pv)、恶性疟原虫(Pf)、三日疟原虫(Pm)和卵形疟原虫(Po)等6种12型病原体的悬液芯片检测方法。方法依据国家生物技术信息中心(NCBI)公开数据库中上述病原体的基因序列信息,设计并合成相关引物及探针,建立多重聚合酶链反应体系(多重PCR),产物与核酸探针微球组杂交后于Bio Plex200检测荧光信号值。结果建立的悬液芯片筛查方法具有较高的特异性和敏感性,能对6种12型病原体进行特异的检测。敏感性试验结果表明,Pm、CHIKV、Pv、WNV、DENV-1检测敏感性约为9DNA拷贝(3×10-7ng);Po、Pf、JEV、DENV-2、DENV-3、DENV-4和YFV检测敏感性约为90DNA拷贝(3×10-6 ng)。结论本研究建立的多重PCR技术结合悬液芯片筛查方法能快速、敏感、特异地同时检测6种共12型蚊媒传染病病原体,为疾病诊断及媒介生物携带病原体快速筛查和鉴定提供了新的手段。     

Arbovirus infections and viral haemorrhagic fevers in Uganda: a serological survey in Karamoja district, 1984

Sera collected in May 1984 from 132 adult residents of Karamoja district, Uganda, were examined by haemagglutination inhibition tests for antibodies against selected arboviruses, namely Chikungunya and Semliki Forest alphaviruses (Togaviridae); dengue type 2, Wesselsbron, West Nile, yellow fever and Zika flaviviruses (Flaviviridae); Bunyamwera, Ilesha and Tahyna bunyaviruses (Bunyaviridae); and Sicilian sandfly fever phlebovirus (Bunyaviridae); and by immunofluorescence tests against certain haemorrhagic fever viruses, Lassa fever arenavirus (Arenaviridae), Ebola-Sudan, Ebola-Za?re and Marburg filoviruses (Filoviridae), Crimean-Congo haemorrhagic fever nairovirus and Rift Valley fever phlebovirus (Bunyaviridae). Antibodies against Chikungunya virus were the most prevalent (47%), followed by flavivirus antibodies (16%), which were probably due mainly to West Nile virus. No evidence of yellow fever or dengue virus circulation was observed. A few individuals had antibodies against Crimean-Congo haemorrhagic fever, Lassa, Ebola and Marburg viruses, suggesting that these viruses all circulate in the area.     

Potential arbovirus emergence and implications for the United Kingdom

Arboviruses have evolved a number of strategies to survive environmental challenges. This review examines the factors that may determine arbovirus emergence, provides examples of arboviruses that have emerged into new habitats, reviews the arbovirus situation in western Europe in detail, discusses potential arthropod vectors, and attempts to predict the risk for arbovirus emergence in the United Kingdom. We conclude that climate change is probably the most important requirement for the emergence of arthropodborne diseases such as dengue fever, yellow fever, Rift Valley fever, Japanese encephalitis, Crimean-Congo hemorrhagic fever, bluetongue, and African horse sickness in the United Kingdom. While other arboviruses, such as West Nile virus, Sindbis virus, Tahyna virus, and Louping ill virus, apparently circulate in the United Kingdom, they do not appear to present an imminent threat to humans or animals.     

Arbovirus prevalence in mosquitoes, Kenya

Few studies have investigated the many mosquito species that harbor arboviruses in Kenya. During the 2006-2007 Rift Valley fever outbreak in North Eastern Province, Kenya, exophilic mosquitoes were collected from homesteads within 2 affected areas: Gumarey (rural) and Sogan-Godud (urban). Mosquitoes (n = 920) were pooled by trap location and tested for Rift Valley fever virus and West Nile virus. The most common mosquitoes trapped belonged to the genus Culex (75%). Of 105 mosquito pools tested, 22% were positive for Rift Valley fever virus, 18% were positive for West Nile virus, and 3% were positive for both. Estimated mosquito minimum infection rates did not differ between locations. Our data demonstrate the local abundance of mosquitoes that could propagate arboviral infections in Kenya and the high prevalence of vector arbovirus positivity during a Rift Valley fever outbreak.     

2013年南宁口岸5例输入性登革热监测结果分析

目的 分析2013年南宁口岸回国人员中发现的5例输入性登革热病例的流行病学特征,为出入境登革热监测和制定防治对策提供科学依据.方法 对2013年南宁口岸回国人员体检中发现的5例登革热病例资料进行分析,同时采用ELISA法和实时荧光RT-PCR两种方法进行检测.结果 5例病例均为输入性实验室确诊病例,来自登革热疫区,出现乏力、发热、头痛等临床症状,4例患者的DEV-IgM呈阳性,1例呈阴性,5例的登革病毒实时荧光RT-PCR结果均呈阳性.结论 荧光RT-PCR具有较高的敏感性,适用于登革热早期诊断.近几年南宁市报告的输入性登革热病例增多,应加强进出口岸及边境地区的监测和防控,避免出现因输入病例引起的本地感染或局部暴发疫情.     

海南省虫媒病毒调查研究状况

海南省地处热带北缘,自然条件适合节肢动物繁殖及虫媒病毒传播,曾发生过多次登革热大流行,也是流行性乙型脑炎(乙脑)地方性流行区.以往从海南省多种媒介和宿主动物中分离到4种甲病毒(基孔肯雅、马雅罗、盖塔和罗斯河病毒)、2种黄病毒(乙脑和登革热病毒)和1种Seadornavirus.现对海南省新分离虫媒病毒及其人畜感染状况进行回顾整理,为当地以及我国虫媒病毒病的预防控制提供依据.     

8种重大烈性传染病病毒液相芯片检测方法的建立

目的建立基于多重PCR技术结合液相芯片技术同时检测汉坦病毒（HTV）、裂谷热病毒（RVFV）、黄热病毒（YFV）、西尼罗病毒（WNV）、克里米亚-刚果出血热病毒（CCHFV）、拉沙热[下同]病毒（LFV）、埃博拉病毒（EBV）和马尔堡病毒（MBV）的方法。方法建立8种病毒同时扩增的多重PCR反应体系,分别用各种病毒特异的核酸探针偶联不同编码的微球,将获得的PCR产物与偶联核酸探针的微球混合物进行杂交,建立液相芯片检测方法,并对建立的液相芯片检测方法进行灵敏度及特异性检测评价。结果建立的8种重大烈性传染病病毒的液相芯片筛查方法具有较高的特异性和敏感性,能对8种病毒进行特异的检测。灵敏性实验结果表明,RVFV为10 ng/PCR、WNV为1 ng/PCR、EBV为10 ng/PCR、CCHFV为10 pg/PCR、MBV为1 ng/PCR、HTV为100 pg/PCR、LFV为1 ng/PCR、YFV为10 pg/PCR。结论本研究建立的病毒液相芯片筛查方法能快速、敏感、特异地同时检测8种重大烈性传染病病毒,对口岸入境人员是否携带重大烈性传染病病毒的快速筛查具有广阔的应用前景。     

West nile virus disease and other arboviral diseases - United States, 2011

Arthropodborne viruses (arboviruses) are transmitted to humans primarily through the bites of infected mosquitoes and ticks. Symptomatic infections most often manifest as a systemic febrile illness and, less commonly, as neuroinvasive disease (e.g., meningitis, encephalitis, or acute flaccid paralysis). West Nile virus (WNV) is the leading cause of domestically acquired arboviral disease in the United States. However, several other arboviruses also cause seasonal outbreaks and sporadic cases. In 2011, CDC received reports of 871 cases of nationally notifiable arboviral diseases (excluding dengue); etiological agents included WNV (712 cases), La Crosse virus (LACV) (130), Powassan virus (POWV) (16), St. Louis encephalitis virus (SLEV) (six), Eastern equine encephalitis virus (EEEV) (four), and Jamestown Canyon virus (JCV) (three). Of these, 624 (72%) were classified as neuroinvasive disease, for a national incidence of 0.20 per 100,000 population. WNV and other arboviruses continue to cause focal outbreaks and severe illness in substantial numbers of persons in the United States.     

Climate change in spain and risk of infectious and parasitic diseases transmitted by arthropods and rodents

Due to Spain's being located near Africa, being a stopping-off point for migrating birds and individuals and due to its climate conditions, nearing those of areas where there are vector-borne diseases, this is a country where this type of diseases could taken on greater importance due to the climate change. The possible risk would result from the geographical spread of already established vectors or due to subtropical vectors adapted to surviving in cooler, dried climates being imported and taking up residence. Hypothetically, the vector-borne diseases subject to be influenced by the climate change in Spain would be those transmitted by dipterans, such as dengue fever, West Nile encephalitis, Rift Valley fever, malaria and leishmaniasis; tick-transmitted diseases, such as Crimean-Congo hemorrhagic fever, tick-borne encephalitis, Lyme disease, spotted fever and endemic relapsing fever; and rodent-transmitted diseases. But the greatest, most viable threat would be the Aedes albopictus mosquito, which would be capable of transmitting viral diseases such as West Nile encephalitis or dengue fever, taking up residence. But, for actual areas of endemia being established, a combination of other factors, such as the massive, simultaneous influx of animal or human reservoirs and the deterioration of the social healthcare conditions and of the Public Health services.     

Arbovirus infections in Sarawak,October 1968–February 1970: Human serological studies in a Land Dyak Village

449 human sera collected in a Land Dyak village were tested for antibodies to 11 arboviruses. Japanese encephalitis and dengue virus antibodies were particularly prevalent. The rates of infection with these viruses were estimated to be 5 · 2% per annum for Japanese encephalitis, 8 · 8% for dengue 1 and 4 · 3% for dengue 2. Chikungunya virus antibodies were quite common with an annual infection rate of the order of 5% per annum. Infections with other Group A and B and Bunyamwera group viruses were generally at a low level.