一株狂犬病病毒街毒株的分离鉴定与组织培养

对来自河北省无极县某地区疑似狂犬病病犬脑组织进行FAT、电镜观察、RT-PCR扩增,以及MIT法等确诊为狂犬病,对分离到的这株狂犬病街毒命名为WJ07-1,分析N蛋白和G蛋白序列,主要抗原位点发生轻度变异,同源性分析发现N基因与ZhejiangWz(H)株同源性最高为99.0%,G基因与JSL27株同源性最高为99.4%。对分离到的狂犬病病毒进行组织培养发现这株狂犬病病毒街毒株对细胞的适应性和感染性很弱,需要进行多次传代培养,才可以适应N2a、BHK-21细胞。     

狂犬病的研究现状

本文从狂犬病病毒、狂犬病的发病机体及狂犬病的免疫学三方面对狂犬病的研究现状作出综述,希望能为以后狂犬病的研究提供文献依据。     

奶牛狂犬病病例诊断报告

泰安周边某奶牛场先后有8头奶牛发病,临床症状疑似狂犬病症状。采集死亡牛小脑、大脑皮层、大脑海马回等部位的脑组织进行常规切片染色。结果表明,在小脑、大脑海马角神经细胞的胞浆内发现嗜酸性包涵体,证实该奶牛发病为狂犬病毒感染。     

四川省家犬狂犬病免疫抗体监测

为了解四川省各市州犬只的狂犬病抗体水平,本试验采集了四川省南充市、绵阳市、广元市等14个市州的犬只血清样(共计258份),利用荷兰EVL公司的酶联免疫吸附试剂盒进行狂犬病血清抗体检测。结果表明,14个市州的犬只抗体阳性率在20%~50%之间,其中有3个市州的血清阳性率为0。     

腺病毒载体狂犬病疫苗研究进展

狂犬病是一种古老的人兽共患传染病。世界上已有部分国家和地区成功地消灭了狂犬病。在我国,每年有约3 000人死于狂犬病。腺病毒作为载体的优点包括高效的入核机制、优良的转染性、较低的病原性、较高的基因表达量及清楚的基因背景。目前,腺病毒已广泛应用于基础研究、基因治疗和疫苗研发。20世纪末,以腺病毒为载体的表达狂犬病病毒糖蛋白的疫苗就已诞生,一直发展至今。以腺病毒载体狂犬病疫苗作为口服疫苗,可望在控制和消除狂犬病中发挥着巨大的作用。     

Fighting Rabies in Eastern Europe,the Middle East and Central Asia ‐ Experts Call for a Regional Initiative for Rabies Elimination

MEEREB is an informal network of rabies experts from the Middle East, Eastern Europe and Central Asia, seeking to eliminate rabies from the region. They met for the second time to review the current rabies situation, both globally and in their respective countries, highlighting current rabies control problems and potential solutions. Success stories in Latin America, in Western Europe, in some Asian countries, as well as in Croatia and Serbia prove that elimination of human rabies is achievable in the MEEREB region. It requires political willingness and cooperation of all stakeholders, including Ministries of Health and of Agriculture; adequate management of animal bites through post‐exposure prophylaxis; pre‐exposure prophylaxis for populations at high risk of rabies exposure, animal vaccination and humane control of stray dog populations. MEEREB members called for a regional initiative for rabies elimination in Eastern Europe and the Middle East. They are confident that the elimination of human rabies of canine origin can be achieved in the region through adopting a One Health approach, and that campaigns for rabies elimination will have significant benefit for public health, including strengthening the structure for control of other zoonoses.     

狂犬病实验室诊断研究进展

狂犬病是世界上最可怕的人兽共患传染病,所有温血动物对狂犬病均易感,狂犬病已成为世界严重的公共卫生问题。暴露后疫苗接种可以达到有效的预防,狂犬病的预防和暴露后处理措施的有效实施均有赖于实验室的安全、准确、快速诊断。另外,免疫后及时对血清中和抗体效价进行监测,也是防控狂犬病的关键措施。文章从狂犬病实验室诊断的安全问题、实验室检测样本的选择、组织学检查、病毒抗原检测、生物学鉴定、核酸检测及血清抗体检测等方面对狂犬病实验室诊断方法及研究进展等进行综述和评价。     

狂犬病病毒全基因组进化研究进展

狂犬病病毒基因进化研究能够揭示病毒流行病学特点和规律,为有效防控狂犬病提供科学依据.狂犬病病毒基因组5个基因中,目前对N基因和G基因研究比较多,N基因序列分析可用于基因分型,G基因序列分析可用于病毒免疫原性、毒力、宿主转换等相关研究.多株狂犬病病毒已完成全基因组测序,但全基因组水平的进化研究还很不足,因为基因组中的各个基因之间的相互作用,各个基因面临的选择压力不同,单个基因的进化不一定能代表病毒的进化,所以需要不断探索合适的方法对狂犬病病毒全基因组进行进化研究.     

抗狂犬病毒单克隆抗体的纯化及标记

对一株稳定、特异、高效的单克隆抗体（mAb）进行了纯化和荧光素（FITC）的标记,利用标记产物与已有的抗狂犬病毒核蛋白单克隆抗体标记物相比较,效果良好,二者可以结合（或单独）使用;应用标记好的荧光抗体与FITC-抗狂犬病毒免疫球蛋白比较进行血清样品效价测定实验,来确定纯化、标记后的单克隆抗体的灵敏度,结果表明本实验制备的单克隆抗体与FITC-抗狂犬病毒免疫球蛋白具有相同的灵敏度,该单克隆抗体测得的血清样品效价结果与FITC-抗狂犬病毒免疫球蛋白测得的结果相同。因此,此荧光抗体可应用于狂犬病病毒检测（FAT）、病毒毒力测定（TCID50）、中和抗体实验（FAVN）等方面的检测实验,此荧光抗体可为狂犬病的相关研究提供有力的保障。     

Epidemiological Characteristics of Human and Animal Rabies in Azerbaijan

The Caucasus is a region of geopolitical importance, in the gateway between Europe and Asia. This geographical location makes the region equally important in the epidemiology and control of transboundary infectious diseases such as rabies. Azerbaijan is the largest country in the Caucasus, and although rabies is notifiable and considered endemic, there is little information on the burden of human and animal rabies. Here, we describe a cross‐disciplinary international collaboration aimed at improving rabies control in Azerbaijan. Partial nucleoprotein gene sequences were obtained from animal rabies cases for comparison with those from surrounding areas. Reported human and animal rabies cases between 2000 and 2010 were also reviewed and analysed by region and year. Comparison of rabies virus strains circulating in Azerbaijan demonstrates more than one lineage of rabies virus circulating concurrently in Azerbaijan and illustrates the need for further sample collection and characterization. Officially reported rabies data showed an increase in human and animal rabies cases, and an increase in animal bites requiring provision of post‐exposure prophylaxis, since 2006. This is despite apparently consistent levels of dog vaccination and culling of stray dogs.     

狂犬病毒Flury株糖蛋白cDNA克隆与序列分析

根据狂犬病毒糖蛋白核苷酸序列,利用Oliga软件设计两对特异性引物,通过反转录-聚合酶链式反应（RT-PCR）扩增了狂犬病毒Flury-lep糖蛋白的全长cDNA,将其插入克隆载体pMD-18T并测序。测序结果及同源性分析表明,糖蛋白cDNA长1574bp,编码524个氨基酸。狂犬病毒株Flury-lep的RGP基因序列与GenBank公布的狂犬病毒株RGP基因片段核苷酸序列的同源性为82．3％～96．3％,其编码产物的氨基酸序列同源性为88．4％～94．3％。     

Prevalence and serum protein values of strangles (Streptococcus equi) affected mules at Remount Depot,Sargodha (Pakistan)

The prevalence of Streptococcus equi serovar equi (S.equi) in nasal discharge and pus samples from sub‐mandibular lymph nodes in mules at the Remount Depot, Sargodha was examined and total serum proteins, serum albumin, serum globulin and fibrinogen measured. A total of 250 nasal swabs and pus samples were collected from mules and examined microbiologically: 99 (39.6%) were positive for S. equi. A higher occurrence of S. equi was recorded in foals as compared to adults. The concentrations of total serum protein, serum globulin and fibrinogen were significantly increased (P<0.05), while the concentration of serum albumin significantly decreased (P<0.05) in strangles‐affected mules. It was concluded that increased total serum proteins, serum globulin and fibrinogen along with decreased serum albumin were important indicators of infection by S. equi in mules.     

Factors Associated with Dog Rabies Vaccination in Bhol,Philippines: Results of a Cross‐Sectional Cluster Survey Conducted Following the Island‐Wide Rabies Elimination Campaign

The Philippines has a long history of rabies control efforts in their dog populations; however, long‐term success of such programmes and the goal of rabies elimination have not yet been realized. The Bohol Rabies Prevention and Elimination Program was developed as an innovative approach to canine rabies control in 2007. The objective of this study was to assess canine rabies vaccination coverage in the owned‐dog population in Bohol and to describe factors associated with rabies vaccination 2 years after implementation of the programme. We utilized a cross‐sectional cluster survey based on the World Health Organization’s Expanded Programme on Immunization coverage survey technique. We sampled 460 households and collected data on 539 dogs residing within these households. Seventy‐seven per cent of surveyed households reported owning at least one dog. The human‐to‐dog ratio was approximately 4 : 1, and the mean number of dogs owned per household was 1.6. Based on this ratio, we calculated an owned‐dog population of almost 300 000. Overall, 71% of dogs were reported as having been vaccinated for rabies at some time in their lives; however, only 64% of dogs were reported as having been recently vaccinated. Dogs in our study were young (median age = 24 months). The odds of vaccination increased with increasing age. Dogs aged 12–23 months had 4.6 times the odds of vaccination compared to dogs aged 3–11 months (95% CI 1.8–12.0; P = 0.002). Confinement of the dog both day and night was also associated with increased odds of vaccination (OR = 2.1; 95% CI 0.9–4.9; P = 0.07), and this result approached statistical significance. While the programme is on track to meet its goal of 80% vaccination coverage, educational efforts should focus on the need to confine dogs and vaccinate young dogs.     

从克隆的cDNA拯救狂犬病毒HEP-Flury株

为研究狂犬病毒基因的结构,将狂犬病毒HEP-Flury株全基因组cDNA与N、P、L、G4个基因的辅助质粒共转染BHK-21细胞,于体积分数为5％CO2培养箱中培养6d,以狂犬病毒N蛋白荧光抗体染色。结果显示,细胞中含有大量的狂犬病毒粒子,表明已建立了狂犬病毒的反向遗传操作系统。     

应用电镜技术对狂犬病不同毒株的形态学研究

将狂犬病固定毒ＣＴＮ株（中国济南株）、ＣＶＳ株（国际标准攻击毒株）、ＥＲＡ株（兽用疫苗弱毒株）,分别在小鼠脑组织增殖及ＥＲＡ毒株在ＢＨＫ２１传代细胞增殖,通过超薄切片和负染色制备样品,电镜观察。结果发现,在相同鼠脑组织中,ＣＴＮ株病毒粒子比较难查找,而ＣＶＳ株和ＥＲＡ株较易被发现。另外,在不同毒株的脑组织和细胞培养物另还发现了大量的非典型粒子,诸如削了顶的粒子以及锥形、柱形等形态,同时还发现了胞浆包涵体。通过试验,我们认为：⑴典型病毒数量与免疫原性呈正相关。⑵在观察鼠脑细胞中的病毒时,发现有神经髓鞘部位是难以找到病毒粒子的。⑶狂犬病病毒粒子存在于宿主细胞浆内,核内未发现。     

Rabies Vaccination: Higher Failure Rates in Imported Dogs than in those Vaccinated in Italy

The current European Union (EU) legislation decrees that pets entering the EU from a rabies‐infected third country have to obtain a satisfactory virus‐neutralizing antibody level, while those moving within the EU require only rabies vaccination as the risk of moving a rabid pet within the EU is considered negligible. A number of factors driving individual variations in dog vaccine response have been previously reported, including a high rate of vaccine failure in puppies, especially those subject to commercial transport. A total of 21 001 observations collected from dogs (2006–2012) vaccinated in compliance with the current EU regulations were statistically analysed to assess the effect of different risk factors related to rabies vaccine efficacy. Within this framework, we were able to compare the vaccination failure rate in a group of dogs entering the Italian border from EU and non‐EU countries to those vaccinated in Italy prior to international travel. Our analysis identified that cross‐breeds and two breed categories showed high vaccine success rates, while Beagles and Boxers were the least likely to show a successful response to vaccination (88.82% and 90.32%, respectively). Our analysis revealed diverse performances among the commercially available vaccines, in terms of serological peak windows, and marked differences according to geographical area. Of note, we found a higher vaccine failure rate in imported dogs (13.15%) than in those vaccinated in Italy (5.89%). Our findings suggest that the choice of vaccine may influence the likelihood of an animal achieving a protective serological level and that time from vaccination to sampling should be considered when interpreting serological results. A higher vaccine failure in imported compared to Italian dogs highlights the key role that border controls still have in assessing the full compliance of pet movements with EU legislation to minimize the risk of rabies being reintroduced into a disease‐free area.     

Evaluation of Mass Vaccination Campaign Coverage Against Rabies in Dogs in Tunisia

In Tunisia, rabies continues to be considered as a serious public health concern. Very costly mass vaccination of dogs against rabies and expensive post‐exposure prophylaxis are prerequisites to maintain a low level of human rabies cases. In Tunisia, the implementation of mass vaccination campaigns at the national level has undoubtedly contributed to the drop of rabies endemicity, but the overall outcome is rather suboptimal. In this investigation, we wanted to estimate the extent of the vaccination coverage in dogs in three Governorates (Manouba, Kassrine and Mednine), by collecting data through questionnaires and interviews relevant to 1470 owned dogs. When the campaign is correctly applied, as in Manouba, almost all the targeted dog population can be reached by parenteral vaccination and an almost elimination of the disease can be evidenced. However, in Kasserine and especially in Medenine, where the vaccination coverage is lower than 31%, a reservoir for the disease can be maintained. To match the official figures of vaccination coverage, we should update the statistics of the size of dog population which seems to be bigger than what is assumed. In addition, we wanted to assess the level of involvement of the local population with the vaccination campaigns by marking vaccinated dogs with necklaces and establishing later on the vaccination coverage accordingly. The highest levels of vaccination coverage can be reached in the rural regions. Therefore, the low vaccination coverage in rural areas, reported at the national level, is more attributable to the lack of human and financial resources to reach remote regions. We think that rabies control programmes in Tunisia can be successful if vaccination coverage can reach 70% in all parts of the country. The achieved vaccination coverage should be estimated after random visits in many parts of the country and by checking valid vaccination certificates.     

狂犬病病毒致神经功能异常机制的研究进展

狂犬病是一种以神经系统感染为特征的高度致死性人兽共患病毒性传染病,危害大,但其致病机制尚不明确.狂犬病病毒(RV)G蛋白与神经细胞表面受体发生特异性结合决定其嗜神经特性,其他RV结构蛋白对RV感染具有保护和调节作用;机体感染RV后神经元结构和机体内与功能相关的部分蛋白及基因表达水平发生改变,而且RV在神经核的分布可能影...