犬狂犬病病毒ELISA抗体检测试剂盒的研制及初步应用     

廖园园  谢红玲 《中国兽药杂志》2013,47(8):1-4

用狂犬病毒单克隆抗体预包被酶标板,将纯化的狂犬病毒作为检测用抗原,利用捕获包被法建立了间接ELISA法用于犬狂犬病病毒抗体的检测。通过优化ELISA条件,研制试剂盒并应用于犬狂犬疫苗的免疫效果检验。利用研制的试剂盒与RFFIT、商品化同类试剂盒对30份犬血清进行平行检测,总符合率分别为90%和93.33%,与其它几种常见犬病毒抗体阳性血清无交叉反应。试剂盒批内、批间变异系数分别为1.45%～5.79%和2.35%～7.21%,2～8℃保存12个月稳定。基于用单抗预包被后再包被检测抗原,本试剂盒检测样品抗体的灵敏度和稳定性得以显著提高,因此适合于不同来源狂犬疫苗人工免疫犬血清的RV抗体水平监测。  相似文献   

Molecular epidemiological study of animal rabies in Kazakhstan          下载免费PDF全文

Gulzhan N. YESSEMBEKOVA  XIAO Shuang  Assem ABENOV  Talgat KARIBAEV  Alexandr SHEVTSOV  Amirgazin ASYLULAN  Yersyn Y. MUKHANBETKALIYEV  SHUAI Lei  BU Zhi-gao  Sarsenbay K. ABDRAKHMANOV 《农业科学学报》2023,22(4):1266-1275

Rabies is a serious public health issue in Kazakhstan, with high economic impact and social burden.  As part of a routine surveillance, 31 rabies-positive brain specimens taken from livestock (cattle) and carnivores (dogs, foxes, and cats) during 2013–2021 were subject to viral sequencing.  Phylogenetic and Bayesian analysis were performed using obtained rabies virus (RABV) sequences.  All 31 strains of RABV candidate belonged to the Cosmopolitan clade, of which 30 strains belonged to steppe-type subclade, and 1 dog strain belonged to Other subclade.  The 31 strains did not diverge from RABV strains in Kazakhstan and neighboring countries, including Russia, Mongolia, and China, suggesting that animal rabies has close relationship and transmission between borders.  Fox-originated strains and cattle strains shared similar sequence signature, and some animal rabies cases had space–time intersection, showing that infected foxes were a major transmission source of cattle rabies in different Kazakhstan regions.  Besides, free-roaming dogs played a pivotal role in rabies epizootics of cattle in Kazakhstan.  The recent spread of animal rabies presents an increasing threat to public health, and provides updated information for improving current control and prevention strategies at the source for Kazakhstan and neighboring countries.  相似文献   

Evaluating the public health response to a mass bat exposure—Wyoming, 2017     

Andrea Cote  Ryan M. Wallace  David A. Jackson  Maria A. Said  Karl Musgrave  Cuc H. Tran  Clayton Van Houten  Alexia Harrist  Danielle Buttke  Ashley Busacker  Veronica Pickens  Sarah Anne J. Guagliardo 《Zoonoses and public health》2019,66(5):504-511

Mass bat exposures (MBEs) occur when multiple people are exposed to a bat or a bat colony, often over an extended period. In August 2017, a public health investigation was started in response to an MBE that occurred during May–August 2017 at a national park research station in Wyoming. We identified 176 people who had slept primarily in two lodges (Lodges A and B) at the research station, and successfully contacted 165 (93.8%) of these individuals. Risk assessments (RAs) were administered to all 165 individuals to determine degree and type of exposures to bats (e.g., biting or scratching). Exposure status for research station guests was classified as “non‐exposed,” “low risk” or “high risk,” and counselling was provided to guide post‐exposure prophylaxis (PEP) recommendations. Prior to public health notification and intervention, 19 persons made the decision to pursue PEP. The healthcare‐seeking behaviours of this group were taken to represent outcomes in the absence of public health intervention. (These persons received a RA, and their risk classification was retrospectively assigned.) Approximately 1–2 weeks after conducting the RAs, we conducted a follow‐up survey to determine whether recommendations regarding PEP were ultimately followed. The proportion of individuals that unnecessarily pursued PEP was higher among the 19 individuals that sought health care prior to receiving the RA (p < 0.00001). Among those receiving the RA first, all persons classified as high risk followed public health guidance to seek PEP treatment. Despite this, upon re‐interview, only 21/79 (26.6%) of guests could accurately recall their risk classification, with most people (55.7%) overestimating their risk. Study findings demonstrate that early public health interventions such as RAs can reduce unnecessary use of PEP and that messaging used during rabies counselling should be clear.  相似文献   

玉林市犬狂犬病流行状况调查     

甘一波  段振华  黄宇  张红云  刘香林  黄荣春 《畜牧与兽医》2019,(5):131-134

通过调查问卷方式得知2010-2016年玉林市犬只饲养量年均20.13万头,免疫犬年均为13.64万头,免疫密度年均为67.74%,仍存在未经免疫的犬。ELISA方法检测免疫犬抗体阳性率为91.57%(1509/1648),非免疫犬为4.13%(10/242);RT-PCR方法检测520份犬脑组织,病原检出率为5.00%(26/520)。通过数据分析与实际情况结合,表明玉林市犬狂犬病免疫抗体合格率较高,疫苗效果表现稳定,外观健康犬仍携带狂犬病病毒。  相似文献   

Dog ecology and its implications for rabies control in Gwagwalada,Federal Capital Territory,Abuja, Nigeria          下载免费PDF全文

P. P. Mshelbwala  D. K. Akinwolemiwa  B.V. Maikai  R. G. Otolorin  N. A. Maurice  J. S. Weese 《Zoonoses and public health》2018,65(1):168-176

The objectives of this study were to determine the characteristics of a dog population, including their accessibility to vaccination and health care, in urban and semiurban areas of Gwagwalada, Abuja, Nigeria. Direct street counts and a house‐to‐house survey of city streets were performed. A total of 451 households were surveyed comprising 43.7% urban and 53.3% semiurban areas. A total of 848 owned dogs were identified, along with 3,115 corresponding humans. With a dog‐to‐human ratio of 1:3.7, the dog population in the study area was estimated as 103,758. A total of 396 dogs were counted on the streets with the greater proportion (74%) in semiurban areas. Most dogs in semiurban areas (77.3%) had no certificate confirming vaccination against rabies, compared to 47.2% in urban areas (p = .004). The majority of dogs in the urban (60.9%) and semiurban (82.0%) were free roaming. In the multivariable model, age, presence of a collar, region, sex, use and having ever visited a veterinarian were significantly associated with rabies vaccination. The majority (125/197, 63.5%) of respondents with higher education were willing to pay more for the healthcare needs of their dogs as opposed to those with a lower level of education (93/251, 37.1%, p = .001). The study revealed a high dog population density, vaccination coverage below WHO recommendation of 70% and generally reduced healthcare‐seeking behaviour among dog owners in Gwagwalada, Abuja, Nigeria.  相似文献   

狂犬病病毒分子流行病学研究进展   总被引：1，自引：0，他引：1  

赵云蛟  钱爱东  李公美  姚纪元 《经济动物学报》2004,8(3):178-180

介绍了狂犬病病毒的流行特点、基因型和检测方法 ,综述了狂犬病病毒核蛋白和糖蛋白基因的研究进展 ,并简要概述了我国学者对狂犬病病毒N基因和G基因的序列分析比较结果 ,表明我国狂犬病病毒的街毒存在着地域差异。  相似文献   

狂犬病毒CVS株G基因和N基因共表达重组腺病毒载体的构建   总被引：1，自引：0，他引：1  

李江涛  殷相平  张金卫  丁农  柳纪省 《浙江农业学报》2011,23(3):0

通过引入内部核糖体进入位点序列,构建狂犬病毒G基因和N基因共表达的腺病毒载体。通过RT-PCR方法扩增得到RVG基因、N基因。N基因先亚克隆入pIRES质粒;G基因经Kpn I和Mlu I,pIRES-N质粒经Mlu I、Not I双酶切,回收目的基因后与经Kpn I和Not I双酶切处理的腺病毒穿梭载体pAdTrack-CMV连接,再与pAdEasy-1质粒在BJ5183菌中同源重组产生腺病毒载体质粒。线性化后的重组腺病毒质粒转染293细胞,通过观察报告基因绿色荧光蛋白的表达鉴定重组的腺病毒,RT-PCR 法检测狂犬病毒糖蛋白和核蛋白基因片段。该重组病毒质粒经酶切鉴定与预期结果一致;转染293细胞后观察到绿色荧光蛋白的表达;RT-PCR 法可检测到糖蛋白和核蛋白基因片段。试验成功构建了G和N双基因共表达重组腺病毒载体,为狂犬病活载体疫苗的研制提供了依据。  相似文献   

表达狂犬病毒糖蛋白和核蛋白的重组腺病毒的生物学特性及免疫研究   总被引：1，自引：0，他引：1  

殷相平  李志勇  李江涛  李宝玉  兰喜  杨彬  李学瑞  柳纪省 《甘肃农业大学学报》2009,44(4)

对狂犬病毒G+N双基因复制缺陷型腺病毒穿梭载体进行转染,以获得融合表过狂犬病毒糖蛋白和核蛋白重组腺病毒,并对其进行生物学特性和免疫学研究.结果表明:将含有狂犬病毒G+N的重组穿梭质粒pAdTrack-CMV/G+N与腺病毒骨架载体pAdEasy-1在大肠杆菌细胞内同源重组,可获得带有目的基因的重组腺病毒载体pAd/G+N,经PacI酶切后转染HEK-293细胞,可获得带有目的基因的重组腺病毒.重组腺病毒在HEK-293细胞中连续传代15代次后,在细胞内的病变时间缩短为20 h以内.经测定,重组腺病毒对HEK-293细胞的TCID50为10-8.0.mL-1,用RT-PCR法可检测到狂犬病毒糖蛋白和核蛋白基因片段;重组腺病毒经口服免疫小白鼠,对狂犬病毒CVS毒株的免疫保护率可达80%.  相似文献   

狂犬病病毒BD-06株基质蛋白的原核表达、纯化及其多克隆抗体的制备   总被引：2，自引：0，他引：2  

赵雪超  张学炜  张守峰  刘晔  米立娟  扈荣良 《中国畜牧兽医》2013,40(4):36-39

本研究旨在获得抗狂犬病病毒M蛋白的多克隆抗体,为进一步研究M蛋白功能提供材料。本试验以狂犬病病毒BD06毒株为模板克隆M基因序列,将其克隆至原核表达载体pET28a;经酶切和测序鉴定,M蛋白基因序列已正确插入表达载体pET28a;以表达载体pET28a-M转化E.coli Rosetta株,并以0.5 mmol/L IPTG诱导,结果表达出27 ku左右的M蛋白;将诱导表达的蛋白质回收纯化,以纯化的M蛋白多次免疫兔子制备多克隆抗体;Western blotting分析和间接免疫荧光分析结果表明,制得的抗体与病毒能够反应,运用制得的多克隆抗体定位了基质蛋白在感染狂犬病病毒的BHK-21细胞中的位置。结果表明成功制得了抗狂犬病病毒M蛋白的多克隆抗体,为研究狂犬病病毒M蛋白功能奠定了基础。  相似文献   

TaqMan探针荧光RT-PCR检测狂犬病病毒     

王振全  罗宝正  薄清如  周昌芳  白鸽  许远靖  王冲  吴殿君 《中国畜牧兽医》2012,39(6):79-82

根据GenBank已公布的狂犬病病毒(rabies virus,RV)核蛋白(N)基因序列设计并合成一对特异性的引物和探针,建立基于TaqMan探针的荧光RT-PCR检测狂犬病病毒方法。对狂犬病疫苗提取核酸后进行RT-PCR扩增,将目的条带切胶回收,克隆测序,重组质粒作为标准阳性对照。对建立的TaqMan探针荧光RT-PCR方法做灵敏度、特异性、重复性及稳定性试验。结果显示,该方法可以达到10拷贝/μL的灵敏度,可以将狂犬病病毒与犬瘟热病毒、犬细小病毒、犬腺病毒、犬冠状病毒和犬副流感病毒分开,方法重复性好,稳定可靠。  相似文献