首页 | 本学科首页   官方微博 | 高级检索  
相似文献
 共查询到20条相似文献,搜索用时 45 毫秒
1.
Vilcek S  Durkovic B  Kolesarova M  Paton DJ 《Preventive veterinary medicine》2005,72(1-2):31-5; discussion 215-9
Genetic typing of bovine viral diarrhoea virus (BVDV) is important for the precise classification of viruses as well as for the development of molecular epidemiology. BVDV isolates were usually typed based on comparison of genomic sequences from the 5'-untranslated region (5'-UTR), N(pro) and E2 region. Recently we have identified 11 genetic groups (subgenotypes) of BVDV-1. Our further experiments confirmed a new subgenotype, BVDV-1k, isolated from cattle in Switzerland. BVDV isolates from India were typed as BVDV-1b whereas BVDV-1c is a predominant subgenotype in Australia. The results of genetic typing of BVDV indicate that distribution of subgenotypes has no relationship to the geographic origin of viral isolates.  相似文献   

2.
Thirteen BVDV isolates collected in four geographic regions of India between 2000 and 2002 were typed in 5'-UTR. To confirm results of genetic typing, selected viruses were also analysed in the N(pro) region. Phylogenetic analysis revealed that all Indian BVDV isolates belong to BVDV-1b (Osloss-like group). Despite a long distance between the farms from which the viruses were isolated there was no correlation between the origin of viral isolates and their position in a phylogenetic tree. Higher genetic similarity of Indian BVDV isolates was observed most probably due to the uncontrolled movement of cattle as well as the uncontrolled use of semen from bulls for breeding of local and farm cattle in different states of India.  相似文献   

3.
Phylogenetic analyses of bovine viral diarrhea virus (BVDV) were performed based on the nucleotide sequences of the 5' untranslated region (5'-UTR) and E2-coding gene. Thirty-six BVDV detected from naturally infected cattle in the northern region of Japan were divided into three genotypes, BVDV1a, BVDV1b and BVDV2, in a 5'-UTR phylogenetic tree. In a phylogenetic tree constructed from the E2-coding gene, BVDV1c was identified and the viruses classified in BVDV1c were included in BVDV1a in the 5'-UTR phylogenetic tree. Moreover, BVDV1a and BVDV1b in the E2-phylogenetic tree clustered closer together than in the 5'-UTR tree. These results suggested that phylogenetic analysis of the E2 gene was more useful for identification of subgenotypes within BVDV1.  相似文献   

4.
A molecular epidemiology study was performed on a selection of 30 rabies-positive brain samples collected between 1994 and 2010 in Slovenia and originating from the red fox (n=19), badger (n=3), cattle (n=3), dog (n=2), cat (n=1), marten (n=1) and horse (n=1). Based on the comparison of 1092 and 672 nucleotide sequences of nucleoprotein (N) and partial glycoprotein (G) gene regions, a low genetic diversity of the circulating strains was detected, but both phylogenetic trees were consistent with the topology where partial nucleoprotein or glycoprotein genes were used. A high sequence identity in the N and G gene to rabies virus isolates from neighbouring countries was found. The Slovenian strains were clearly different from the vaccine strains SAD B19 and SAD Bern, which have been used in Slovenia since 1988.  相似文献   

5.
In the last decade, several studies were performed to characterise bovine viral diarrhoea virus (BVDV) isolates and define genetic groups by genotyping. Much data is now available from GenBank, predominantly sequences from the 5' untranslated region (5'-UTR). In order to find out whether genetic grouping of isolates from different countries could be harmonised, 22 new isolates from five countries were analysed in combination with published sequences. Eighteen of these isolates were typed as BVDV genotype 1 (BVDV-1), and one isolate from Argentina and three isolates from Brazil were typed as BVDV-2. BVDV-1 isolates were clustered into five previously defined genetic groups: BVDV-1a, b, d, e and f. Two isolates from Finland and one from Egypt formed a group which was tentatively labelled as BVDV-1j, since statistical support was low. By using a fragment of the Npro gene for typing, we found that these isolates fall into the same group as a deer strain, and are statistically significant. Some Swiss BVDV strains taken from GenBank were found in a new genetic group which was designated as BVDV-1k. The BVDV-2 isolates included in this study seemed to fall into two genetic groups.  相似文献   

6.

In order to isolate buffaloes herpesvirus 1 (BuHV-1) from latently infected water buffalo (Bubalus bubalis), 16 buffalo heifers were selected from a herd. At first, animals were bled and their sera were tested by virus neutralization (VN) test, using bovine herpesvirus 1 (BoHV-1). According to the results of VN test and dexamethasone injection (0.1 mg/kg BW) for 5 consecutive days, the examined buffaloes were divided into 4 groups. Vaginal and nasal swabs were daily collected from all buffaloes from day 0 to 10 days later. Based on the cytopathic effects in cell culture, a herpesvirus was isolated only from nasal swabs of three seropositive buffaloes which they had received dexamethasone. The nasal swabs of these three buffaloes were also positive in PCR, using primers specific for ruminant herpesviruses gD gene. The identity of the isolated viruses was determined according to partial amino acid sequences of gD, deduced from the nucleotide sequences of the PCR products. On the basis of sequence alignment, phylogenetic analysis, and genetic distances, the three buffalo virus isolates were more closely related to BuHV-1 and BoHV-5 than to BoHV-1.

  相似文献   

7.
8.
9.
为了解近年来云南省师宗县蓝舌病病毒流行情况,2012年在师宗县五龙乡建立了10头蓝舌病血清学阴性黄牛的监控动物群。从2012年5~10月,每周采血1次,11~12月,每月采血1次,采用C-ELISA进行血清学监测。8月开始动物血清学检测结果转阳性,至11月,监控动物全部转为阳性。用转阳前1周、转阳本周、转阳后2~13周的经处理的红细胞静脉接种鸡胚,收获鸡胚肝脏,用PBS悬浮捣碎的鸡胚肝脏,上清接种于C6/36细胞一代、BHK-21三代后,出现细胞病变(cytopathic effect,CPE)。采用RT-PCR方法,针对蓝舌病较为保守的血清型群特异片段VP7设计了2对引物,扩增其相应片段。结果显示,共分离到86份疑似分离物,其中67份疑似分离物细胞培养液上清经RT-PCR扩增,均扩增出1156 bp片段,初步确认为蓝舌病病毒。采用国际24个蓝舌病标准毒及24个标准阳性血清对86份疑似分离物及其对应血清进行细胞微量中和试验,67份毒株为蓝舌病病毒,与RT-PCR结果一致。通过对2份经中和试验定型为BTV-1、BTV-16分离株的VP2基因测序分析发现,BTV-1株序列与同型Y863(登录号:KC879616)参考毒株的同源性为92%,BTV-16株序列与登录号为AB686221的毒株同源性为99%。结果表明共分离到67株蓝舌病毒株,分离株主要为BTV-1、BTV-9、BTV-16三个血清型。  相似文献   

10.
The presence of chicken anemia virus (CAV) in Slovenia was confirmed by inoculation of 1-day-old chickens without antibodies against CAV and isolation of the virus on the Marek's disease chicken cell-MSB1 line and by polymerase chain reaction (PCR). Experimental inoculation of 1-day-old chickens resulted in lower hematocrit values, atrophy of the thymus, and atrophy of bone marrow. CAV was confirmed by PCR in the thymus, bone marrow, bursa of Fabricius, liver, spleen, ileocecal tonsils, duodenum, and proventriculus. The nucleotide sequence of the whole viral protein (VP)1 gene was determined by direct sequencing. Alignment of VP1 nucleotide sequences of Slovenian CAV isolates (CAV-69/00, CAV-469/01, and CAV-130/03) showed 99.4% to 99.9% homology. The VP1 nucleotide sequence alignment of Slovenian isolates with 19 other CAV strains demonstrated 94.4% to 99.4% homology. Slovenian isolates shared highest homology with the BD-3 isolate from Bangladesh. Alignment of the deduced VP1 amino acids showed that the Slovenian isolates shared 100% homology and had an amino acid sequence most similar to the BD-3 strain from Bangladesh (99.6%) and were 99.1% similar to the G6 strain from Japan and the L-028 strain from the United States. The Slovenian isolates were least similar (96.6%) to the 82-2 strain from Japan. A phylogeneric analysis on the basis of the alignment of the VP1 amino acids showed that CAV isolates used in the study formed three groups that indicated the possible existence of genetic groups among CAV strains. The CAV isolates were grouped together independent of their geographic origin and pathogenicity.  相似文献   

11.
Using RNA purified directly from stored clinical specimens, a collection of 62 pestiviruses were typed by RT-PCR and sequencing within the 5'-untranslated region of the genome. All the specimens had been obtained in 1966/1967 from diary cattle in England and Wales. Eight further pestiviruses, grown in cell culture, were characterised in the same way. Seven of these viruses were representatives of a panel of British isolates, obtained from cattle ten years before. The eighth was the virus used in a British bovine viral diarrhoea (BVD) vaccine. Most of the viruses were genetically unique and were of BVDV type Ia. One recent isolate was BVDV type Ib, two others were intermediate between Ia and Ib. No BVDV type II or border disease virus (BDV) isolates were found. There was no overall association between geographical and phylogenetic clustering, suggesting long-distance virus dispersal, presumably via trading of infected cattle. The sequences of the recently obtained cattle viruses were very similar or, in one case, identical to the older isolates in the region studied. Their close similarity to some previously characterised pestiviruses from British sheep suggests that a common pool of BVDV Ia is shared by these two livestock species, although another pestivirus--BVDV--is confined to sheep. The British cattle viruses were mostly distinct from continental European isolates, but more similar to type Ia isolates from North American cattle.  相似文献   

12.
The prevalence of bovine viral diarrhoea virus (BVDV) biotypes and subgenotypes was determined from 131 BVDV positive samples from a diagnostic laboratory. The majority of the isolates were from Oklahoma; however, other states including Kansas, Texas, and Arkansas were represented. These BVDV samples were from submissions of 76 live animals and 55 necropsy samples. There were 131 BVDV samples represented by 117 noncytopathic (NCP), 11 cytopathic (CP) and 3 cases with mixed NCP and CP biotypes. The NCP isolates were more common (P < 0.05) than the CP and NCP/CP combination. The BVDV samples were segregated into three subgenotypes by differential PCR and sequencing of a viral genomic region, 5'-untranslated region (5'-UTR). There were more BVDV1b subgenotypes 60/131 (45.8%) than BVDV1a, 37/131 (28.2%) or BVDV2a, 34/131 (26.0%) (P < 0.05). The organ system involvement included the major categories such as respiratory, digestive, mixed/multiple organs, abortions, and persistent infections (PI). All three BVDV subgenotypes were found in persistently infected (PI) cattle and respiratory diseases, both major requests for BVDV diagnosis. Only one of the 131 viruses was genetically similar to the strains present in U.S. vaccines.  相似文献   

13.
14.
Genetic heterogeneity of Turkish ruminant pestiviruses was investigated by phylogenetic analysis of complete N(pro) encoding nucleotide sequences. A total of 30 virus isolates obtained from 15 provinces around the country between 1997 and 2005 were included in the phylogenetic analysis. Virus isolates mostly originated from cattle with one isolate from sheep. The bovine isolates all belonged to BVDV-1, the sheep isolate to BVDV-2. Fifteen isolates formed a new subgroup within BVDV-1, tentatively named BVDV-1l. The remaining bovine isolates were typed as BVDV-1a (n=4), BVDV-1b (n=4), BVDV-1d (n=3), BVDV-1f (n=2) and BVDV-1h (n=1). The isolates allocated to BVDV-1l originated from various geographical regions in different years. There was no correlation between genetic grouping and locations where isolates were obtained. Viruses originating from one farm in most cases belonged to the same subgroup (n=5). This study indicates that the newly detected subgroup BVDV-1l is predominant and widespread in Turkey. Moreover, an ovine virus isolate was identified as the first member of BVDV-2 reported in Turkey. A serological survey using samples from western Turkey indicated that BVDV-2 is also present in cattle.  相似文献   

15.
16.
Reported here are the results of antigenic and genetic characterisation of equine influenza strains causing local outbreaks reported to the Equine Diagnostic Centre in Berlin, Germany. In 2000, equine influenza virus was detected in a nasal swab from a non-vaccinated horse using a rapid diagnostic kit, but was not successfully isolated. Partial direct sequencing of the haemagglutinin (HA1) gene, indicated that the virus was a European lineage H3N8 subtype strain representative of strains isolated in several European countries during 2000. In 2002, two equine influenza viruses were isolated from nasal swabs both taken from unvaccinated horses with acute respiratory symptoms housed at the same stables. Antigenic characterisation using a panel of ferret antisera suggested that these isolates also belonged to the European lineage of H3N8 viruses. Analysis of deduced HA1 amino acid sequences confirmed that the HA1 of both isolates were identical and belonged to the European lineage. However, from phylogenetic analysis, both strains appeared to be more closely related to viruses isolated between 1989 and 1995 than to viruses isolated more recently in Europe. These results suggested that viruses with fewer changes than those on the main evolutionary lineage may continue to circulate. The importance of expanding current equine influenza surveillance efforts is emphasised.  相似文献   

17.
Eighty-nine bovine viral diarrhoea viruses (BVDV) from Australia have been genetically typed by sequencing of the 5' untranslated region (5'-UTR) and for selected isolates the N(pro) region of the viral genome. Phylogenetic reconstructions indicated that all of the samples examined clustered within the BVDV type 1 genotype. Of the 11 previously described genetic groups of BVDV-1, 87 of the samples examined in this study clustered with the BVDV-1c, while two samples clustered with the BVDV-1a. Based on these analyses there appears to be limited genetic variation within the Australian BVDV field isolates. In addition, the phylogenetic reconstructions indicate that the clustering of Australian BVDV in the phylogenetic trees is not a result of geographic isolation.  相似文献   

18.
Twenty nine recent isolates of bovine viral diarrhea-mucosal disease (BVD-MD) virus, 17 from persistently infected cattle and 12 from mucosal disease, were compared antigenically with the reference strains by a serum neutralization test. The reference viruses were divided into 2 groups, tentatively designated as N and K, based on the antigenic relationships in the cross-neutralization test. Antigenic properties of recent isolates were considerably different with the sources of virus isolation. Seventeen isolates recovered from persistently infected cattle were divided into 3 groups in the neutralization test using antisera to the reference strains; 12 and 2 were considered as the possible members of groups K and N, respectively, and the others belonged to neither group. On the other hand, 10 of 12 isolates recovered from mucosal disease were considered as the possible members of group N, and the others were classified into neither group. Interestingly, none of BVD-MD viruses isolated from cases of mucosal disease belonged to group K. The results of serologic survey on sera collected from 713 cattle at the Hokkaido provinces in 1974 to 1988 indicated that infections of cattle with BVD-MD viruses other than group K were prominent before 1981. Cattle infected with group K BVD-MD virus were first detected in 1982, and increased in number thereafter. The results obtained in this study suggested that BVD-MD viruses with various antigenic properties spread widely among cattle herds, and also a possibility that clinical manifestations in cattle infected with BVD-MD viruses may differ with their antigenic properties.  相似文献   

19.
猪流感病毒H1N1、H1N2和H3N2亚型多重RT-PCR诊断方法的建立   总被引:2,自引:3,他引:2  
对我国分离到的猪流感病毒和GenBank数据库中已有的猪流感病毒H1N1、H1N2和H3N2亚型毒株的HA、NA基因核苷酸序列进行分析,分别选出各个病毒亚型HA和NA基因中高度保守且特异的核苷酸区域,设计扩增猪流感病毒H1和H3、N1和N2亚型的2套多重PCR特异性引物,建立了猪流感H1N1、H1N2和H3N2亚型病毒多重RT-PCR诊断方法。采用该方法对H1N1、H1N2、H3N2亚型猪流感病毒标准参考株进行RT-PCR检测,结果均呈阳性,对扩增得到的片段进行序列测定和BLAST比较,表明为目的基因片段。其它几种常见猪病病毒和其它亚型猪流感病毒的RT-PCR扩增结果都呈阴性。对107EID50/0.1mL病毒进行稀释,提取RNA进行敏感性试验,RT-PCR最少可检测到102EID50的病毒量核酸。对40份阳性临床样品的检测结果是H1N1、H1N2和H3N2亚型分别为16份、1份和20份,其它3份样品同时含有H1N1和H3N2亚型猪流感病毒,和鸡胚分离病毒结果100%一致。试验证明建立的猪流感病毒H1N1、H1N2和H3N2亚型多重RT-PCR诊断方法是一种特异敏感的诊断方法,可用于临床样品的早期快速诊断和分型。  相似文献   

20.
1株牛病毒性腹泻病毒分离毒株的基因组特征   总被引:3,自引:3,他引:0  
旨在从宁夏某奶牛群持续感染牛分离牛源牛病毒性腹泻病毒(BVDV),并解析其基因组特征,为研究我国不同地区BVDV分离株遗传演化规律提供理论依据。利用BVDV抗原检测试剂盒检测宁夏回族自治区银川市某示范区的240头高产奶牛间隔两周的双份抗凝血,筛选持续感染牛,分离血液淋巴细胞制备裂解液接种牛肾细胞(MDBK),分离鉴定获得BVDV株,克隆测序获得全基因组序列,比较分析其遗传演化关系。从该示范区高产奶牛筛选获得2头持续感染牛,分离获得1株非致细胞病变型BVDV,命名为NX2019/01。测序获得基因组全序列(12 107 nt),其中ORF长11 703 nt,编码3 898个氨基酸。在基因组水平,NX2019/01株与我国SD-15、ZM-95、XC、LN-1等1m亚型分离株相似性较高(92.17%~93.84%),但ErnsE1以及E2基因存在较大差异。示范区同群牛急性感染BVDV时,毒株E2蛋白N端编码区核苷酸突变可导致第9位或第67位氨基酸变异。重组分析表明,NX2019/01株E2基因179—288位核苷酸区段以及ZM-95株E1基因168位—E2基因332位核苷酸区段存在相似的重组信号,可能由主要亲本SD-15株与次要亲本LN-1株重组形成,表明NX2019/01株、ZM-95株在演化进程中与SD-15株以及LN-1株或早期流行的高度相似毒株存在密切关联。本研究从持续感染高产奶牛分离获得了牛源BVDV-1m亚型毒株,在基因组水平厘清了BVDV-1m亚型毒株的进化关系,并首次发现同亚型BVDV毒株基因同源重组,为进一步研究BVDV在我国的演化规律奠定了基础。  相似文献   

设为首页 | 免责声明 | 关于勤云 | 加入收藏

Copyright©北京勤云科技发展有限公司  京ICP备09084417号