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The Diagnosis of Rabies in a Horse by Brain Neutralization Test   总被引:1,自引:1,他引:0       下载免费PDF全文
A horse showing clinical signs of a neurological disorder was killed and various diagnostic tests for rabies were carried out. Histopathlogy revealed a nonsuppurative encephalitis. Fluorescent antibody test and mouse inoculation test were negative. A positive diagnosis of rabies was based on a high antibody titer (1:10,000) to rabies virus in brain tissue.  相似文献   

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Brain tissue from 64 rabies suspect specimens were inoculated intracerebrally into twenty 9-12 gm adult Swiss white mice. Two mice from each specimen were killed on specific days postinoculation and examined for the presence of rabies virus by the fluorescent antibody staining technique. In this way a positive diagnosis was made in the majority of cases between postinoculation days 4 and 12 when the incubation period of these same specimens ranged between eight and 20 days.  相似文献   

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Inclusion bodies, indistinguishable from rabies inclusion bodies (Negri bodies), were found in the brains of 8 nonrabid dogs. The inclusions were compared to Negri bodies present in neurons of rabies-positive animals and examined for the presence of rabies virus by a combination of immunoperoxidase staining (7 cases), fluorescent antibody (FA) staining (1 case), and transmission electron microscopy (4 cases). Positive immunoperoxidase staining for rabies was obtained in brain tissues from FA rabies-positive animals. All brain tissues from the 7 dogs stained by the immunoperoxidase method and the brain from the 1 dog stained by the FA method were negative for rabies. Rabies virus was not found in inclusion-containing neurons in the cases examined by transmission electron microscopy. These results emphasize the importance of FA testing and mouse inoculation for the diagnosis of rabies.  相似文献   

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A tissue culture infection test in routine rabies diagnosis.   总被引:3,自引:0,他引:3       下载免费PDF全文
A cell culture infection test was developed for the isolation of rabies virus from field cases submitted for rabies diagnosis. The procedure involved the addition of a suspension of suspect brain tissue to a suspension of murine neuroblastoma cells in 96-well microtiter plates. The cultures were then incubated at 35-36 degrees C for four days at which time they were fixed, stained with a fluorescein-labelled hamster antirabies antibody conjugate and examined with a fluorescence microscope. Rabies antigen in cells was readily visible as brilliant, apple-green fluorescent particles. This technique was compared with the standard mouse inoculation test and was at least as sensitive to infection with small amounts of virus, required a much shorter test period and was substantially more economical than the mouse inoculation test. The new cell culture test is now in use at this laboratory, replacing the mouse inoculation test.  相似文献   

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Four cats were inoculated IM with rabies virus isolated from the salivary gland of a naturally infected big brown bat (Eptesicus fuscus). The 4 cats developed clinical signs of rabies after a median incubation period of 42 days. The median duration of clinical illness was 5 days. Results of fluorescent antibody evaluation, mouse inoculation, and tissue culture isolation indicated large differences in virus concentrations in various areas of the CNS of individual cats. These differences also were observed between cats. Rabies virus was isolated from the salivary glands and saliva of 2 cats; urinary bladder was the only other nonneural tissue found infected. Our observations indicated that cat rabies can be caused by bat rabies virus; that cats thus infected have infectious saliva during aggressive behavior and can therefore transmit the disease; and that adequate specimens of hippocampus, cerebellum, and brain stem are essential for reliable determination of rabies infection. The findings support recommendations for regular rabies vaccination of cats, even in areas of rabies-free terrestrial mammals.  相似文献   

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Rabies is one of the oldest known zoonotic diseases that has significant impact on public health, but still remains neglected in Serbia. Rabies virus can infect humans and other mammals and causes inflammation of the brain associated with encephalomyelitis and neurological symptoms. In 2010, Veterinary Directorate (national Competent Authority for animal health in Serbia) has started multi‐annual project of oral rabies vaccination of foxes and other wild carnivores (e.g. jackals), as support of long‐term programme of eradication of rabies in Serbia, co‐funded by EU (financed by Instrument for Pre‐Accession Assistance). Monitoring of the effectiveness of oral vaccination campaigns has been carried out in continuation from 2011 and was based on: (i) post‐mortem laboratory examination of brain tissue of target animals (foxes, jackals and other carnivores) by fluorescent antibody test (FAT), (ii) detection of antibodies against rabies virus in serum samples by ELISA and (iii) detection of tetracycline biomarker in the mandibles for the evaluation of vaccine bait uptake. From September 2011 to May 2014, the total number of 4943 brain tissue samples, 4241 sera and 4971 mandibles were analysed. Confirmed rabies‐positive brains decreased from 10 in 2011/2012 to 6 in 2012/2013 and eventually to 1 positive case in 2013/2014. The seroconversion rate increased from 10.48% (133/1269) in 2011/2012 to 20.11% (362/1800) in 2012/2013 and 42.23% (495/1172) in 2013/2014. Along with the seroconversion, the number of detected tetracycline‐positive mandibles demonstrated an increasing tendency in the same period, being 49.67% (682/1373) in 2011/2012, 62.60% (1294/2067) in 2012/2013 and 90.33% (1383/1531) in the monitoring programme carried out in 2013/2014. Presented results confirmed that ORV of foxes and other wildlife in Serbia against rabies was successful and characterized by steady increase of vaccine baits uptake and immunization of animals.  相似文献   

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以灭活的狂犬病病毒CVS株细胞毒免疫BALB/c小鼠,通过间接ELISA法和Western-blot筛选获得针对磷蛋白的单克隆抗体4株:1C9、4B10、2G12、4G5,其中1C9针对氨基端保守表位。以亲和层析法纯化1C9单抗腹水,异硫氰酸荧光黄标记制备荧光抗体。以1C9磷蛋白荧光抗体与本实验室研制的狂犬病核蛋白免疫荧光抗原检测试剂盒,对本实验室收集的501份疑似狂犬病鼬獾、蝙蝠、犬和黄鼬的脑组织样品进行直接免疫荧光平行检测。结果显示,两种检测手段对基因1型狂犬病毒的检出结果完全一致,而蝙蝠源Irkut病毒仅能以磷蛋白单抗1C9检出。本研究成功获得了与我国现有不同基因型狂犬病毒良好反应的抗狂犬病磷蛋白单抗,并应用于狂犬病的直接免疫荧光检测,为狂犬病诊断提供了敏感性和可靠性良好的诊断试剂。  相似文献   

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European bat lyssavirus type 1 (EBLV-1, genotype 5) is known to endemically circulate in insectivorous bat populations in Germany. In August 2001, a rabies suspect stone marten (Martes foina) was found in the city of Burg (Saxony-Anhalt, Germany) and was sent to the regional veterinary laboratory for routine rabies diagnosis. Whereas brain samples repeatedly tested negative in the fluorescent antibody test for classical rabies virus (genotype 1), the mouse inoculation test and the rabies tissue culture inoculation test yielded positive results. Rabies viral RNA was also detected in the stone marten brain sample both by nested and heminested RT-PCR specific for the nucleoprotein gene and for the nucleoprotein phosphoprotein junction of rabies virus. The amplification products were sequenced to genotype the isolate. Sequence data obtained from the first-round RT-PCR products were analysed and the suspect stone marten isolate was confirmed as a rabies related virus (EBLV-1a). Phylogenetic comparison with sequences from recent genotype five isolates from Germany and Denmark showed that it was closely related to a previous isolate of EBLV-1 from a serotine bat in Saxony-Anhalt obtained in the same year in an area adjacent to the place where the EBLV-1 infected stone marten was found. Both EBLV-1 isolates share a 99.5% identity. This is the first report of an EBLV-1a spill-over from an insectivorous bat into wildlife in Europe.  相似文献   

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Three strains of rabies virus, one fixed and two street, were compared for invasiveness in neurectomized rats following foot pad inoculation. Approximately one-fifth of test rats studied revealed that the virus bypassed the severed sciatic and saphenous nerves and reached the brain.

All rats dying within 20 days were positive for rabies upon fluorescent antibody and mouse mortality tests.

Viremia was demonstrated in 20 per cent of both neurectomized or non-neurectomized rats one hour after inoculation of rabies virus, both fixed and street strains, into the foot pad.

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Rabies is caused by several Lyssavirus species, a group of negative sense RNA viruses. Although rabies is preventable, it is often neglected particularly in developing countries in the face of many competing public and veterinary health priorities. Epidemiological information based on laboratory-based surveillance data is critical to adequately strategise control and prevention plans. In this regard the fluorescent antibody test for rabies virus antigen in brain tissues is still considered the basic requirement for laboratory confirmation of animal cases. Occasionally brain tissues from suspected rabid animals are still submitted in formalin, although this has been discouraged for a number of years. Immunohistochemical testing or a modified fluorescent antibody technique can be performed on such samples. However, this method is cumbersome and cannot distinguish between different Lyssavirus species. Owing to RNA degradation in formalin-fixed tissues, conventional RT-PCR methodologies have also been proven to be unreliable. This report is concerned with a rabies case in a domestic dog from an area in South Africa where rabies is not common. Typing of the virus involved was therefore important, but the only available sample was submitted as a formalin-fixed specimen. A real-time RT-PCR method was therefore applied and it was possible to confirm rabies and obtain phylogenetic information that indicated a close relationship between this virus and the canid rabies virus variants from another province (KwaZulu-Natal) in South Africa.  相似文献   

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Rabies is a zoonotic, fatal and progressive neurological infection caused by rabies virus of the genus Lyssavirus and family Rhabdoviridae. It affects all warm-blooded animals and the disease is prevalent throughout the world and endemic in many countries except in Islands like Australia and Antarctica. Over 60,000 peoples die every year due to rabies, while approximately 15 million people receive rabies post-exposure prophylaxis (PEP) annually. Bite of rabid animals and saliva of infected host are mainly responsible for transmission and wildlife like raccoons, skunks, bats and foxes are main reservoirs for rabies. The incubation period is highly variable from 2 weeks to 6 years (avg. 2–3 months). Though severe neurologic signs and fatal outcome, neuropathological lesions are relatively mild. Rabies virus exploits various mechanisms to evade the host immune responses. Being a major zoonosis, precise and rapid diagnosis is important for early treatment and effective prevention and control measures. Traditional rapid Seller's staining and histopathological methods are still in use for diagnosis of rabies. Direct immunofluoroscent test (dFAT) is gold standard test and most commonly recommended for diagnosis of rabies in fresh brain tissues of dogs by both OIE and WHO. Mouse inoculation test (MIT) and polymerase chain reaction (PCR) are superior and used for routine diagnosis. Vaccination with live attenuated or inactivated viruses, DNA and recombinant vaccines can be done in endemic areas. This review describes in detail about epidemiology, transmission, pathogenesis, advances in diagnosis, vaccination and therapeutic approaches along with appropriate prevention and control strategies.  相似文献   

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European bat lyssavirus type 1 (EBLV‐1, genotype 5) is known to endemically circulate in insectivorous bat populations in Germany. In August 2001, a rabies suspect stone marten (Martes foina) was found in the city of Burg (Saxony‐Anhalt, Germany) and was sent to the regional veterinary laboratory for routine rabies diagnosis. Whereas brain samples repeatedly tested negative in the fluorescent antibody test for classical rabies virus (genotype 1), the mouse inoculation test and the rabies tissue culture inoculation test yielded positive results. Rabies viral RNA was also detected in the stone marten brain sample both by nested and heminested RT‐PCR specific for the nucleoprotein gene and for the nucleoprotein phosphoprotein junction of rabies virus. The amplification products were sequenced to genotype the isolate. Sequence data obtained from the first‐round RT‐PCR products were analysed and the suspect stone marten isolate was confirmed as a rabies related virus (EBLV‐1a). Phylogenetic comparison with sequences from recent genotype five isolates from Germany and Denmark showed that it was closely related to a previous isolate of EBLV‐1 from a serotine bat in Saxony‐Anhalt obtained in the same year in an area adjacent to the place where the EBLV‐1 infected stone marten was found. Both EBLV‐1 isolates share a 99.5% identity. This is the first report of an EBLV‐1a spill‐over from an insectivorous bat into wildlife in Europe.  相似文献   

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对湖南部分地域的8个县(市、区)的8个城区、乡镇和农村的健康犬抽样采集脑组织标本142份,应用RT-PCR和直接荧光抗体试验(FAT)检测,检测结果142份犬脑样中均不含狂犬病病毒。调查上述监测点及周边近三年狂犬病的免疫情况,免疫率均超过70%。表明湖南部分犬免疫工作较好的地区,通过加大群免疫密度,可以使犬狂犬病病毒携带率降低,这可能与形成免疫屏障有关。  相似文献   

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OverviewRabies virus belongs to the genus Lyssavirus, together with European bat lyssaviruses 1 and 2. In clinical practice, rabies virus is easily inactivated by detergent-based disinfectants.InfectionRabid animals are the only source of infection. Virus is shed in the saliva some days before the onset of clinical signs and transmitted through a bite or a scratch to the skin or mucous membranes. The average incubation period in cats is 2 months, but may vary from 2 weeks to several months, or even years.Disease signsAny unexplained aggressive behaviour or sudden behavioural change in cats must be considered suspicious. Two disease manifestations have been identified in cats: the furious and the dumb form. Death occurs after a clinical course of 1–10 days.DiagnosisA definitive rabies diagnosis is obtained by post-mortem laboratory investigation. However, serological tests are used for post-vaccinal control, especially in the context of international movements.Disease managementPost-exposure vaccination of cats depends on the national public health regulations, and is forbidden in many countries.Vaccination recommendationsA single rabies vaccination induces a long-lasting immunity. Kittens should be vaccinated at 12–16 weeks of age to avoid interference from maternally derived antibodies and revaccinated 1 year later. Although some vaccines protect against virulent rabies virus challenge for 3 years or more, national or local legislation may call for annual boosters.  相似文献   

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目前狂犬病疫苗的效力检验采用NIH法,需要使用狂犬病毒强毒株进行攻毒试验,具有一定的生物安全风险。为建立检测小鼠血清抗体效价的间接ELISA方法,代替NIH法中的脑内攻毒试验,扩增狂犬病毒(RV)G蛋白基因,并将其克隆至大肠杆菌pET-32a载体上进行表达。以该蛋白作包被抗原,摸索试验条件,建立检测小鼠血清抗体效价的间接ELISA方法。使用此方法与国际公认的荧光抗体病毒中和试验(FAVN)法比较,两者检测结果曲线相关系数为0.986,表明两者相关性较好,但ELSIA方法更加快捷、简便。本试验建立的间接ELISA方法可用于检测小鼠血清中狂犬病抗体,为狂犬病疫苗效力检验替代方法的建立提供了基础。  相似文献   

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对带有绵羊MT启动子-狂犬病病毒糖蛋白基因的(MT-Rgp)的TgN(oMT-Rgp)2Lge、TgN(oMT-Rgp)4Lge和TgN(oMT-Rgp)6Lge3株转基因小鼠系(子代)进行了表达检测。ELISA结果表明,小鼠肝脏、肾脏中均有糖蛋白表达产物,以肝脏的表达量较高;对肝脏的免疫组化检测结果显示,糖蛋白分布在肝脏实质细胞,主要位于细胞膜上和胞浆内。对8只TgN(oMT-Rgp)2Lge子代鼠一次接种灭活的狂犬病病毒8202株,接种前和接种后3周的血清抗体水平变化在转基因鼠和非转基因鼠之间有明显区别,非转基因鼠抗狂犬病病毒抗体水平明显升高,而转基因鼠抗体水平基本保持不变。经统计学分析,转基因鼠的抗体水平变化相差不显著,表明转基因小鼠对该病毒形成了部分免疫耐受性。  相似文献   

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