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1.
为了评估猪基因Ⅰ型乙型脑炎病毒YA1株、CZ株灭活疫苗的免疫原性,通过将YA1株、CZ株乙脑病毒分别在乳鼠脑内连续传代,获得了高病毒含量的鼠脑病毒液。将佐剂ISA206分别与灭活的YA1株、CZ株乙脑病毒液混合,制备成鼠脑灭活苗。将YA1株、CZ株鼠脑灭活苗以及HW1株商品化鼠脑灭活苗分别免疫接种小鼠、豚鼠、仔猪,采用间接ELISA试验、血凝抑制试验、病毒中和试验以及攻毒保护试验分别检测IgG抗体、血凝抑制抗体、中和抗体的的产生情况以及疫苗对小鼠的攻毒保护率。结果显示YA1株、CZ株灭活苗在小鼠和仔猪上产生的IgG抗体、在豚鼠上产生的血凝抑制抗体、在仔猪上产生的中和抗体以及对小鼠的攻毒保护率均高于HW1株。结果表明乙脑病毒YA1株、CZ株灭活苗的免疫原性优于HW1株,其中CZ株灭活苗的免疫原性最优。  相似文献   

2.
猪乙型脑炎减毒活疫苗毒株的选育研究   总被引:3,自引:0,他引:3  
以乙脑弱毒SA14-14-2作为母种,采用代地鼠肾细胞和乳鼠皮下传代方法并结合蚀斑纯化技术,选育获得乙脑减毒活疫苗猪用毒株SA14-14-2VS株。该毒株经细胞传到15-18代,脑内接种12-14g小鼠不引起发病和死亡;皮下接种10-12g小鼠,从脑组织中不能分离出病毒;经3-5日龄乳鼠回传一代后,脑内毒力Log LD50仅为1.32-2.10,皮下接种小鼠无致病力。该毒株细胞18代毒,用乳猪传至第五代,从乳猪血液、脑、肝组织中分离的病毒再经细胞传二代,其滴度与原毒液相近,并仍对小鼠脑内感染不致死;8头不公猪经该毒接种后,未发生睾丸炎,睾丸组织中未回收到病毒;经静脉接种的2头怀孕30d的初产母猪,蝇接种后2d和4d血样中检出病毒,但接种21d剖杀时,胎儿均健尖,胎盘、羊水及胎儿脑组织中均未回收到病毒,该毒株能使豚鼠和猪产生较强的免疫应答,对小白鼠攻击的保护力比灭活疫苗主;纯毒及外源因子污染 结果表明,该毒株是纯将的乙脑病毒,符合兽用生物制品种毒标准。  相似文献   

3.
为弄清上海地区活禽批发市场中H9禽流感病毒(Avian influenza virus,AIV)的流行情况及鸡群的免疫情况,2009年对上海三大活禽批发市场进行了采样监测。采用HI试验检测H9 AIV抗体、荧光RT-PCR试验和鸡胚接种分离鉴定病毒。共采集110批次1 646份血样和喉头泄殖腔棉拭样品,平均抗体合格率为60.27%,分离到H9病毒134株,其中4-6月和9-11月为全年中病毒分离的2个高峰期(样品带毒率均超过了10.00%),明显比其它月份要高(其他月份均低于5.00%),样品带毒率平均为8.14%。不同市场、不同地区采集的样品其抗体合格率和样品的带毒率也存在一定的差异。在30批分离到病毒的样品中,13批次已免疫H9N2油乳剂灭活苗且抗体合格率均大于70.00%的样品中分离到45株病毒(45/195),其中6批次抗体合格率达到100%的样品中也分离到了病毒(8/90),但带毒率明显比未经疫苗免疫的样品(79/255)低。调查结果表明养殖户对肉鸡群H9N2油乳剂灭活苗免疫重视程度不够,鸡群中带毒现象较普遍。疫苗免疫后能产生较高的免疫抗体,且抗体能减轻临床症状,降低带毒率,但不能完全阻止病毒复制,存在高抗体下带毒现象。  相似文献   

4.
猪繁殖呼吸综合征病毒S3毒株的分离及其免疫特性研究   总被引:2,自引:0,他引:2  
从某猪场仔猪体内分离到一株PPRSV毒株,该毒株能在Marc-145细胞上增殖产生致细胞病变,该病变能被PPRSV美洲型阳性血清所抑制,不被乙脑病毒、猪细小病毒、伪狂犬病毒阳性血清所抑制,经美洲型PPRSV荧光抗体染色呈阳性反应,电镜观察到直径55-60nm的病毒粒子,接种阴性仔猪未见临床症状异常,但抗体检测阳性。命名该分离毒为PPRSV-S3毒株。在S3弱毒株分离鉴定的基础上,进一步在猪体上对其致病性和免疫力进行研究。S3毒株接种仔猪后,仔猪不表现任何症状,也不向外排毒。4-5周龄的仔猪接种S3株后可产生坚强的免疫力,免疫后4-6个月能抵抗强毒攻击。后备母猪配种前2-3周免疫接种S3株,怀孕90-95d攻强毒,未发生流产、死胎、木乃伊胎、产弱仔等繁殖障碍现象。结果表明,S3是一株自然分离的弱毒株,且具有良好的安全性和免疫力。  相似文献   

5.
从福州某鸭场大批发病和死亡的15~20日龄雏鸭肝脾中分离到1株病毒,该病毒无血凝性,可使4日龄健康鸭90.6%发病、62.5%死亡,利用Ⅰ型鸭肝炎病毒(DH)标准血清进行鸡胚中和试验和血清被动免疫保护试验,证明该分离的病毒株为I型DHV。  相似文献   

6.
绵羊慢病毒北疆株自然感染新疆美利奴羊的研究   总被引:3,自引:0,他引:3  
选12只自然感染绵羊慢病毒的成年新疆美利奴羊,用琼脂凝胶免疫扩散试验连续检查血清中OvLV沉淀抗体,发现抗体应答的类型发生转换,即只出现对病毒核心蛋白抗原的抗体,出现对病毒包膜糖蛋白的抗体,同时出现对p25和pg135二者的抗体互相转换。从试验羊的多个器官中分离到3株OvLV,分别命名为OvLV NXJ-55,NXJ-73和NXJ-0418株,其TCID50/mL值诊次为1×10^-5.6,1×1  相似文献   

7.
应用Vero-E6细胞从患狂犬病毒死亡的梅花鹿脑组织中分离出一株狂犬病病毒,经免疫荧光检查,能与狂犬病阳性血清特异性结合。其与人狂犬病阳性血清的结合能被抗狂犬病马血清阻断,所分离毒能通过0.45μm滤膜,超薄切片电镜观察,病毒颗粒清晰可见,为长柱状,Vero-E6细胞培养病毒不致细胞变,在病毒接种第2天,细胞感染率在50%以上,第5天可达90%。  相似文献   

8.
对鸡新城疫病毒V4株血凝效价,感染力,毒力,耐热性以及不同接种途径免疫鸡只的血清HI抗体水平和攻毒保护率进行了测定,证明该株病毒的血凝效价和感染滴度都较高,并且无毒,耐热,接种后可使免疫鸡只对强毒的攻击产生坚强的抵抗力。唯以任何途径免疫均不能诱导接种鸡产生高的血清HI抗体水平。  相似文献   

9.
H3亚型猪流感病毒分离与鉴定   总被引:4,自引:1,他引:4  
从东莞和鹤山等地不同猪场采集的40份鼻拭子或病死猪的肺、气管病料中分离到4株有血凝活性的病毒,其中3株病毒与新城疫病毒阳性血清的HI试验为阳性,另外1株病毒与抗猪流感H3亚型猪流感病毒阳性血清的HI试验为阳性;根据猪流感病毒M基因设计引物,扩增出预期的约315 bp片段,表明该病毒为H3亚型猪流感病毒。  相似文献   

10.
通过对黑龙江省7个禽肉厂养禽场的饲养情况,疲劳免疫、消毒隔离措施及疫情发生情况进行调查,统计各禽肉厂新城疫疫情的流行情况,并对出口的18批次90栋舍540只宰前肉用禽类进行新城疫(NDV)血清血凝抑制抗体检测和病毒分离鉴定。共从12批次25栋舍87只肉用禽中分离出新城疫病毒,并对其中69株新城疫病毒进行毒力鉴定。均为中等以下毒力,来分离出强毒株。  相似文献   

11.
云南省乙型脑炎病毒宿主和媒介研究   总被引:18,自引:0,他引:18  
于1978年至1997年,从云南猪血液和乳猪脑组织中分离到乙型脑炎(JE)病毒5株,从蝙蝠脑组织中分得JE病毒10株,从鸟类脑组织中分得JE病毒5株,从15种蚊虫体内分离到JE病毒63株,从2种蠓类中分得JE病毒2株。分析认为,猪是JE病毒的主要扩散宿主,蝙蝠和鸟类在该病毒保存和传播中起重要作用;三带喙库蚊是该病毒的主要传播媒介,伪杂鳞库蚊、霜背库蚊、蚊腿库蚊、白蚊伊和刺扰伊蚊为重要媒介,蠓类亦可  相似文献   

12.
Sera collected from the southern parts of Japan were subjected to serological tests for antibodies to 24 arthropod-borne or suspected arthropod-borne viruses. A high incidence (82%) of hemagglutination-inhibiting (HI) antibodies was found with Japanese encephalitis (JE) virus. HI antibodies to other Flaviviruses, Murray Valley encephalitis, Apoi, Kunjin, Stratford and Kokobera, were also found in some of the sera, but seemed to be due to cross reaction with JE virus. High neutralizing (NT) antibody incidences were obtained with Akabane (60%) and Aino (30%) viruses known to be endemic in Japan. NT antibodies were also found for Bunyaviruses, Batai and Wongla; Reoviridae viruses, D'Aguilar, Warrego, and Mitchell River; and Kowanyama and Belmont viruses. Complement fixing antibodies were found for Reoviridae viruses bluetongue type 1 and Ibaraki; Picornavirus Nodamura and Rhabdovirus bovine ephemeral fever. No antibodies were detected with Reoviridae viruses Corriparta and Eubenangee; Bunyavirus Trubanaman; and Alfavirus Chikungunya.  相似文献   

13.
We report the results of a 6-year serological and virological monitoring performed in ducks and coots in Italy, in order to assess the degree of influenza A virus circulation in these birds during wintering. A total of 1039 sera collected from 1992 to 1998 was screened by a double antibody sandwich blocking ELISA (NP-ELISA): seroprevalence of antibodies to influenza A viruses was significantly higher in ducks compared to coots (52.2% vs. 7.1%, respectively). The hemagglutination-inhibition (HI) assay, performed on NP-ELISA positive sera, showed that 16.9% of these duck sera and 33.3% of these coot sera had antibodies to at least one influenza virus HA subtype: ducks showed HI antibodies against most of the HA subtypes, except for the H3, H4, H7, and H12; coots were seropositive to the H3 and H10 subtypes, only. From 1993 to 1998, 22 virus strains were obtained from 802 cloacal swabs, with an overall virus isolation frequency of 2.7%. Viruses belonging to the H1N1 subtype were by far the most commonly circulating strains (18/22) and were isolated mainly from ducks (17/18). The remaining viruses were representative of the H10N8, H5N2 and H3N8 subtypes. Our data indicate some differences between influenza A virus circulation in sympatric ducks and coots and a significant antigenic diversity between some reference strains and viruses recently isolated in Italy.  相似文献   

14.
为了解牛感染帕利亚姆亚群达圭勒病毒(D’Aguilar virus,DAGV)情况,本研究应用BHK21细胞对2019年云南省景洪市采集的健康黄牛血液样品进行盲传病毒分离,对出现细胞病变的样品进行形态学、基因组带型和分子生物学鉴定,对分离到的病毒进行S2、S3、S7基因序列测定和比对分析。结果显示,有5个血液样品可致BHK21细胞病变,电镜观察到完整病毒颗粒呈球形,直径约50 nm;琼脂糖凝胶电泳结果发现5株新分离病毒基因组为10节段,呈现3-3-4的电泳带型特征,其带型特征与2014年在云南分离到的DAGV V106/YN/2014毒株相似;5株病毒S2、S3、S7基因核苷酸、氨基酸序列相似性均为100%,S3、S7片段核苷酸和氨基酸序列与中国本土分离的帕利亚姆亚群病毒(Palyamserogroup virus,PALV)毒株相似性最高;S2片段氨基酸与核苷酸序列与日本分离的DAGV相似性最高。S2、S3、S7基因遗传进化分析结果显示,5株新分离毒株间基因相似度为100%;5个毒株的S7与S3基因序列均与中国本土分离的已知部分PALV毒株在同一分支,亲缘关系较近,提示这5株毒株为PALV;5株毒株S2片段核苷酸序列与日本分离的部分DAGV毒株位于同一分支上,亲缘关系较近,进一步证实这5株毒株为PALV DAGV。本研究成功分离到5株DAGV毒株,并进行了基因片段遗传进化分析,为进一步开展DAGV流行病学研究提供基础。  相似文献   

15.
【目的】 了解西南边境地区牛感染中山病病毒(Chuzan virus,CHUV)的情况。【方法】 应用BHK21细胞对云南景洪、江城采集的312份健康黄牛血液样品盲传分离病毒,对出现细胞病变的毒株进行形态学、基因组带型和分子生物学鉴定,对分离到的病毒进行S7、S2基因序列测定和比对分析。【结果】 4份血液样品可致BHK21细胞病变,电镜观察病毒颗粒呈球形,直径约50 nm;琼脂糖凝胶电泳发现,4株分离毒株基因组为10节段,带型为"3-3-4",与2012年云南师宗分离的CHUV SZ187毒株相似;4株病毒S7、S2基因核苷酸序列相似性分别为98.7%~99.7%和97.9%~99.1%;S2基因片段核苷酸和氨基酸序列与中国本土分离的SZ187、GHN-GS-16等毒株相似性最高;S7基因片段核苷酸和氨基酸序列与日本分离的ON-1/E/18、ON-3/E/17及中国本土的SZ187、GHN-GS-16等毒株的相似性最高。S7、S2基因遗传进化分析结果显示,4株新分离毒株亲缘关系最近;4株毒株的S7基因序列与中国本土和日本分离的已知帕利亚姆血清群病毒(Palyam serogroup virus,PALV)毒株位于同一分支,提示4株毒株为PALV,且形成一簇,有一定的地域性;4株毒株S2基因片段核苷酸序列与中国本土分离的CHUV位于同一分支,亲缘关系较近,进一步证实这4株毒株为CHUV。【结论】 本研究首次报告了从云南边境地区牛群中分离到CHUV毒株,为CHUV在中国的流行病学和边境地区疫病风险防控提供了重要参考及有力依据。  相似文献   

16.
本研究旨在了解云南省景洪市虫媒病毒的流行情况。2019年在景洪市勐罕镇设立了3头哨兵动物牛,定期采血进行虫媒病毒的分离与鉴定,共获得7株病毒分离物。经病毒核酸的RT-PCR鉴定,分离到2株血清型分别为6型和7型流行性出血热病毒(epizootic haemorrhagic disease virus,EHDV),2株血清型分别为4型和5型的蓝舌病病毒(bluetongue virus,BTV),1株帕利亚血清群病毒(palyam serogroup virus,PALV)中的D’Aguilar virus(DAV)血清型病毒和2株未鉴定出的环状病毒。经病毒Seg-2、Seg-3序列ORF区的比对和进化分析显示,7株病毒的地域型均为Eastern型,与日本、澳大利亚和印度毒株具有最近的亲缘关系。3头哨兵动物的血液和血清,经病毒核酸及血清中和试验检测,证明3头动物均被相应的病毒感染。动物感染病毒后,血清中的特异性抗体迅速上升,3~4周后达最高点并能够在该水平维持较长时间,而血液中病毒核酸含量2~4周到达最高点后则呈迅速下降趋势。本研究报道了景洪虫媒病毒的分离、毒株序列特征以及在动物上的感染特性,研究结果为进一步了解当地的牛虫媒病毒提供数据支撑,同时3头牛分离获得7株病毒,提示当地可能还存在更多种类的虫媒病毒。  相似文献   

17.
The purpose of this study was to understand the prevalence of arboviruses in Jinghong city,Yunnan province.Three sentinel cattle were set up in Menghan township of Jinghong city in 2019,blood samples were collected regularly for isolation and identification of arboviruses,a total of 7 virus isolates were obtained.Viral nucleic acid were identification by RT-PCR,and the results showed that two strains of epidemic hemorrhagic fever virus (EHDV) with serotypes of 6 and 7,two strains of bluetongue virus (BTV) with serotypes 4 and 5,D’Aguilar virus (DAV) serotype in one strain of PALV and two unidentified circoviruses were isolated,respectively.The ORF region of virus Seg-2 and Seg-3 sequences was compared and analyzed,all the 7 regional strains of the virus were Eastern,which was most closely related to strains in Japan,Australia and India.The blood and serum of three sentinel animals were tested by viral nucleic acid and serum neutralization test,which proved that all three animals were infected with the corresponding virus.When animals were infected with the virus,specific antibodies in the serum rise rapidly,it reached peaks 3 to 4 weeks later and could remain at this level for a long time.However,the content of viral nucleic acid in the blood decreased rapidly after reaching the peak from 2 to 4 weeks.The isolation,sequence characteristics and infection characteristics of Jinghong arboviruses in animals were reported in this study,the results provided data support for further understanding of local bovine arboviruses.At the same time,7 strains of virus were isolated from 3 cattle,suggesting that there might be more arboviruses in the area.  相似文献   

18.
Twenty-six captive, adult Egyptian fruit bats (Rousettus aegyptiacus) were tested for the presence of rabies virus neutralizing antibodies (RVNA) using a rapid fluorescent focus inhibition test before and after vaccination. The bats were randomly assigned into three treatment groups: group A (n = 10) bats each received one 0.1-ml dose of monovalent inactivated rabies vaccine, group B (n = 10) bats each received two 0.1-ml doses of vaccine given 30 days apart, and group C (n = 6) bats remained unvaccinated. Plasma was collected from all bats before vaccination and on days 14, 30, 60, and 360. All bats were seronegative before vaccination, and all unvaccinated animals remained negative throughout the study. Rabies virus neutralization titers remained above 0.5 IU/ml from day 30 through day 360 for both vaccinated groups. Group B had significantly higher titers on day 60. This study demonstrated a measurable humoral immune response after vaccination with an inactivated rabies vaccine, with two doses producing a higher level of RVNA. This study confirms the feasibility of a rabies vaccination program for Egyptian fruit bats.  相似文献   

19.
A recombinant baculovirus (RBV) encoding the nucleoprotein (NP) of avian influenza virus (AIV) was generated and the appropriate protein was expressed in Sf9 cells. Purified recombinant NP and the NP-specific monoclonal antibody HB65 were used to establish a competitive ELISA (cELISA) system for the detection of NP-specific antibodies in sera of ducks, geese and wild birds. Tests to evaluate this method were carried out using sera of ducks experimentally infected with AIV, pre-immune duck and chicken sera, and poultry field sera, which tested negative in the haemagglutination inhibition (HI) assay, and field sera of several poultry species experimentally infected with other viruses. The evaluation of the test demonstrated a high sensitivity and specificity of this method. Tests carried out using field sera of duck and goose flocks revealed widely corresponding results obtained by HI assay and cELISA indicating that this test is applicable for flock diagnosis. Differing results were obtained for individual samples. It can be assumed that for the most part this was because of a better recognition of the conserved NP antigen by serum antibodies, although some results remained unclear.  相似文献   

20.
In the period December 1979-May 1980 a respiratory disease spread rapidly through pig herds in The Netherlands. Surveillance of 12 pig farms resulted in isolation of 22 influenza A-Swine-H1N1 (Hsw1N1) strains from 9 pig herds. The morbidity rate was high but the mortality rate was nil. Retardation in growth was observed. Sera collected from affected pig herds showed a fourfold increase in haemagglutination inhibition (HI) titre against A-Swine-H1N1 virus. Sera collected on five farms showed a geometric mean HI titre against the A-H3N2 virus above 100. A significant HI titre increase against this virus was found in sera collected on three farms. These findings indicated a recent infection by this virus. A-H3N2 virus was not isolated. The Dutch Swine-1980 isolates showed in the cross-HI test a distant antigenic relationship with the classical A/Swine/Iowa/30 (H1N1) virus and one-sided close antigenic relationship with A/New Jersey/76 (H1N1) virus.HI antibody to A/Swine/Nederland/80 (H1N1) virus was found in 4, 0, and 44%, to A/New Jersey/76 (H1N1) virus in 0.5, 0.4, and 42%, and to A/Swine/Iowa/30 (H1N1) virus in 0.5, 1, and 30% of pig sera collected in 1976, 1977, and 1980, respectively. HI antibody to A/Hong Kong/68 (H3N2) virus was detected in 36, 56, and 68%, and to A/Victoria/75 (H3N2) virus in 38, 73, and 68% of these sera, respectively.The results of this study indicate that pigs in The Netherlands, like those in North America, Southeast Asia, Japan, and Western Europe harbour A-Swine-H1N1 and A-H3N2 influenza viruses and are thus potential reservoirs for future human pandemics.  相似文献   

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