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1.
《中国兽医杂志》2015,(9)
为研究来自河南省不同猪源的副猪嗜血杆菌(Hps)的遗传演化关系,应用PCR方法扩增所分离的良种猪源、家养野猪源和地方土猪源等3株Hps(KF0901、JZ0801和XY0501)的16S r RNA基因,并进行序列比较分析。结果 3株Hps的16S r RNA基因全长均为822 bp,彼此间核苷酸序列同源性为99.3%~99.6%,与参考菌株的核苷酸序列同源性为97.1%~99.4%;基于16S r RNA基因序列绘制的系统进化树显示,本研究中家养野猪源Hps和地方土猪源Hps均属于血清5型,而良种猪源血清5型Hps却与血清12型和血清14型的亲缘关系更近。表明Hps在良种猪、家养野猪和地方土猪之间彼此交叉感染或具有共同来源;并非所有血清5型Hps分离株都属同一分支。 相似文献
2.
为测定猪源脑心肌炎病毒(EMCV)NJ08分离株全基因序列,本研究应用RT-PCR方法分5个基因片段扩增NJ08分离株的基因组,同时应用3'-RACE和5'-RACE技术扩增3'-UTR和5'-UTR,分别克隆于pMD18-T载体中进行测序,获得EMCV NJ08株全基因组cDNA序列,并将全基因组及其推导的氨基酸序列与GenBank中登录的国内外参考病毒株进行同源性比较及系统进化分析.结果表明:EMCV NJ08分离株基因组全长7 724bp;属于Ia亚群;与GXLC、GX0602、BJC3、HB1等国内分离株以及BEL-2887A、CBNU、K3、K11、EMCV-R、PV21等国外分离株同源性达99%以上;EMCV流行株的非结构蛋白中3D最为保守,2A变异最大,结构蛋白中VP2最为保守,VP1变异最大.本研究丰富了我国EMCV分子流行病学资料. 相似文献
3.
猪源和鼠源脑心肌炎病毒分离株基因组的比较分析 总被引：4，自引：0，他引：4
为了分析猪源和鼠源脑心肌炎病毒(EMCV)基因组的差异,对分离自同一猪场猪临床样本和鼠组织样本的2株病毒(GX0601和GX0602)进行了全基因组序列测定和比较分析.结果显示,2个分离毒株的基因组全长(包含poly A)分别为7 729和7 725 nt,核苷酸和氨基酸同源性均在99.8%以上.与国外毒株及国内已报道的分离株的序列比较结果表明,2个毒株与BJC3和HBl的核苷酸同源性为98.18%～99.41%,推导的氨基酸序列同源性为99.5%～99.7%,与国外分离株的核苷酸序列和氨基睃序列的同源性分别介于80.53%～99.57%和93.1%～99.5%.基于基因组开放阅读框推导的氨基酸序列的系统进化分析表明,我国分离毒株与国外毒株PEC9、CBNU、BEL-2887A/91和EMCV-R的同源性较高,位于同一个亚群.研究结果提示,猪源与鼠源毒株的基因组具有很高的同源性,鼠是猪场EMCV感染的传染来源. 相似文献
4.
猪脑心肌炎病毒GXLC株VP1基因的克隆与序列分析 总被引：4，自引：2，他引：2
应用RT-PCR方法扩增猪脑心肌炎病毒GXLC株的VP1基因,扩增产物克隆入pMD18-T载体后进行测序,对获得的VP1基因序列进行分析。序列分析表明,GXLC株VP1基因长度为831 bp,编码277 aa,含有2个潜在的N-糖基化基序和9个抗原表位。同源性分析表明,GXLC株与国内外其他26个EMCV分离株VP1基因核苷酸序列的同源性在78.6%～99.6%之间、氨基酸序列的同源性在95.1%～99.3%之间。遗传进化分析表明,基于VP1基因序列绘制的系统进化树可以将所有EMCV分离株分成两个群,即Ⅰ群和Ⅱ群,Ⅰ群可再细分为Ⅰa亚群和Ⅰb亚群,其中猪源、鼠源EMCV在Ⅰ群和Ⅱ群中均有分布,人源EMCV则分布在Ⅱ群;GXLC株与其他中国分离株均属于Ⅰa亚群。 相似文献
5.
目前国内对猪轮状病毒的研究较少,作者旨在对猪A群轮状病毒进行分离与鉴定,为后续猪轮状病毒致病机理和分子生物学特性研究奠定基础.用胰蛋白酶处理RT-PCR检测猪A群轮状病毒病原阳性的临床腹泻粪样,然后接种长成单层的MA-104细胞,进行病毒分离传代.再对分离毒株进行常规RT-PCR鉴定和电镜观察,并对分离毒株的VP6、VP7和VP8基因进行测序及序列分析.结果成功分离猪A群轮状病毒TM-a株,经序列同源性分析发现,与TM-a株VP6、VP7和VP8基因同源性最高的毒株分别为中国北京人源LL3354株、印度人源RMC321株和日本野猪源GUB-71株,其相似性为96.0％、95.1％和97.0％.该TM-a株轮状病毒不同基因表现出与人源轮状病毒和猪源轮状病毒的高度同源性,推测猪A群轮状病毒可能在人畜间传播,发生基因重组现象. 相似文献
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《畜牧与兽医》2014,(7):65-68
采用RT-PCR方法对河南省某猪场送检的家养野猪病料进行检测,表明其病原为猪流行性腹泻病毒(PEDV),命名为JZ0801,并对其S1基因克隆后进行序列比较和遗传变异分析。结果显示:JZ0801与PEDV参考毒株的核苷酸序列同源性为86.8%99.0%,与疫苗株CV777的核苷酸同源性仅为87.1%;PEDV可分为Ⅰ、Ⅱ和Ⅲ群,2011年后流行的PEDV主要分布在Ⅰ群和Ⅱ群;JZ0801分布于Ⅰ群,与国内猪源分离株HN6-201211和HBMC2012亲缘关系最近,而与日本、韩国毒株在遗传关系上关系较远。研究结果证实,家养野猪源PEDV与目前我国各地流行PEDV的主要类群相一致;PEDV可感染家养野猪并发病,提醒在进行野生动物养殖活动中要充分考虑疫病传播的生态学;PEDV存在较大的地域差异。 相似文献
7.
猪脑心肌炎病毒的分离与鉴定 总被引：24，自引：0，他引：24
从规模化猪场疑似病例组织病料中分离到5株脑心肌炎病毒（EMCV），对其中的2株（EMCV BJC3和EMCV HB1）进行了系统鉴定。结果表明，病毒粒子大小约为27nm，呈圆形；2株病毒均不耐酸，对氯仿不敏感，对胰蛋白酶敏感性有所不同，60℃加热1h可被灭活，二价阳离子对病毒没有明显的保护作用。用EMCV多克隆抗体作间接免疫荧光，分离毒株感染的BHK-21细胞的胞浆中可见特异性荧光。对分离毒株进行了VP1和3D基因的扩增和序列测定，结果表明，2个毒株VP1基因的核苷酸序列同源性为99．49％，氨基酸序列的同源性为98．46％，与GenBank中EMCV毒株的核苷酸序列同源性介于81．61％～99．59％，氨基酸序列同源性在95．5％以上；3D基因扩增片段序列与其它毒株的核苷酸与氨基酸序列同源性均为100％。基于VP1基因的系统进化树表明，2株分离毒均位于进化树的主干上，变异度不大。由此表明，EMCV感染已在我国猪场存在。 相似文献
8.
猪脑心肌炎病毒GXLC株的分离及其3D基因分子特征的分析 总被引：2，自引：0，他引：2
采集临床疑似脑心肌炎死亡仔猪的组织作为接种材料,接种于BHK-21细胞系,观察细胞病变(CPE),并用RT-PCR和间接荧光抗体试验(IFA)进行鉴定,证实分离到1株脑心肌炎病毒(encephalomyocarditis virus,EMCV),命名为GXLC株。应用RT-PCR方法扩增GXLC株的3D基因,扩增产物克隆入pMD18-T载体后进行测序,对获得的3D基因序列进行分析。序列分析结果表明,GXLC株3D基因全长1380 nt,编码460个氨基酸,含有7个抗原表位。同源性分析结果表明,GXLC株与国内外其它EMCV分离株3D基因核苷酸序列的同源性在84.7%~99.7%之间,氨基酸序列的同源性在96.1%~99.6%之间。遗传进化分析结果表明,基于3D基因核苷酸序列绘制的系统进化树可将所有EMCV分离株分成2个群:Ⅰ群和Ⅱ群,Ⅰ群可再细分为Ⅰa亚群和Ⅰb亚群,其中猪源EMCV在Ⅰ群和Ⅱ群中均有分布,而鼠源EMCV分布在Ⅰ群,人源EMCV分布在Ⅱ群;GXLC株与其它中国分离株均属于Ⅰa亚群。 相似文献
9.
采集临床疑似脑心肌炎死亡仔猪的组织作为接种材料,接种于BHK-21细胞系,观察细胞病变（CPE）,并用RT-PCR和间接荧光抗体试验（IFA）进行鉴定,证实分离到1株脑心肌炎病毒（encephalomyocarditis virus,EMCV）,命名为GXLC株。应用RT-PCR方法扩增GXLC株的3D基因,扩增产物克隆入pMD18-T载体后进行测序,对获得的3D基因序列进行分析。序列分析结果表明,GXLC株3D基因全长1380 nt,编码460个氨基酸,含有7个抗原表位。同源性分析结果表明,GXLC株与国内外其它EMCV分离株3D基因核苷酸序列的同源性在84.7%～99.7%之间,氨基酸序列的同源性在96.1%～99.6%之间。遗传进化分析结果表明,基于3D基因核苷酸序列绘制的系统进化树可将所有EMCV分离株分成2个群：Ⅰ群和Ⅱ群,Ⅰ群可再细分为Ⅰa亚群和Ⅰb亚群,其中猪源EMCV在Ⅰ群和Ⅱ群中均有分布,而鼠源EMCV分布在Ⅰ群,人源EMCV分布在Ⅱ群;GXLC株与其它中国分离株均属于Ⅰa亚群。 相似文献
10.
《畜牧与兽医》2016,(9):32-37
将1份经RT-PCR检测猪轮状病毒阳性的临床腹泻粪样感染Vero细胞,连续盲传8代,出现病变,成功分离到1株病毒,经RT-PCR检验和电镜观察,鉴定该病毒为轮状病毒,命名为GD-01-2015。扩增该病毒的VP4和VP7基因进行分型和系统进化分析,结果发现,其VP4基因为P[7]型,与来自韩国的猪源毒株K71同源性达到99.8%;其VP7基因为G5型,与来自韩国的猪源毒株06-6-1同源性达到99.7%。由此可以推测GD-01-2015株与韩国猪源毒株有相似的进化来源。根据轮状病毒分类委员会(RCWG)提出的A群轮状病毒的最新分类方法,GD-01-2015株基因型为G5P[7]。本研究为监测轮状病毒的流行状况及其疫苗的研制提供了理论依据。 相似文献
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James H.d.T. van Sandwyk Nigel C. Bennett Robert Swanepoel Armanda D.S. Bastos 《Veterinary microbiology》2013
Encephalomyocarditis virus (EMCV) outbreaks are rare in southern Africa. Only two have been reported to date from South Africa, both coinciding with rodent irruptions. The first outbreak manifested as acute myocarditis in pigs in 1979, whilst the second, occurring from 1993 to 1994, was linked to the deaths of 64 free-ranging adult African elephants (Loxodonta africana). The P1 genome region, inclusive of the flanking leader (L) and 2A genes, of three South African isolates, one from swine and two from elephants, was characterised by PCR amplification and sequencing of up to 11 overlapping fragments. In addition to the resulting 3329 nucleotide dataset, the 3D region that is widely used in molecular epidemiology studies, was characterised, and three datasets (P1, VP1/3 and 3D), complemented with available homologous EMCV data, were compiled for analyses. Phylogenetic inferences revealed the near-identical elephant outbreak strains to be most closely related to a mengovirus from rhesus macaques (Macaca mulatta) in Uganda, differing from the latter by between 11% (3D) and 15% (VP3/1). The South African pig isolate differed by 4% (3D) and 11% (VP3/1) from available European and Asian pig virus sequences. This study confirms the presence of two genetically distinct EMCV lineages recovered from sporadic outbreaks in wild and domestic hosts in southern Africa, and provides valuable baseline data for future outbreak eventualities in the sub-region. 相似文献
13.
Lorenzetti E da Silva Medeiros TN Alfieri AF Alfieri AA 《Veterinary microbiology》2011,154(1-2):191-196
Group A rotavirus (RV-A) with short electropherotype was identified by ss-PAGE in a neonatal diarrhea outbreak at a Brazilian pig farm where the sows were regularly vaccinated with a commercial vaccine containing OSU (G5P[7]) and Gottfried (G4P[6]) porcine RV-A (PoRV-A) strains. The ss-PAGE positive stool samples (n=20) were characterized as P[6] genotype by multiplex-nested-RT-PCR assay. The nucleotide analysis of the VP4 gene (VP8*) state that the viruses clustered in P[6] lineages that are also shared by RV-A strains identified in human hosts. Nucleotide analysis of the VP7 gene identified different lineages in G4 including a new lineage tentatively designated IX. The immunological pressure induced by commercial vaccine with a rotavirus containing a G4P[6] genotype of porcine origin (Gottfried strain) might have allowed the selection of PoRV-A strains with characteristics found in RV-A strains isolated of human hosts, such as P[6]-Ie and If, and promoted the selection or emergence of RV-A strains with a new lineage of the G4 genotype. The characterization of PoRV-A strains with unusual genotypes described in this study highlight the importance of surveys on the relationship between human and animal rotavirus strains. 相似文献
14.
Epidemiologic, pathogenic and molecular analysis of recent encephalomyocarditis outbreaks in Belgium. 总被引：4，自引：0，他引：4
F Koenen H Vanderhallen F Castryck C Miry 《Zentralblatt für Veterin?rmedizin. Reihe B. Journal of veterinary medicine. Series B》1999,46(4):217-231
In 1991 EMCV was isolated for the first time in Belgium from the offspring of a sow with reproductive failure. From August 1995 until December 1996, EMCV was diagnosed in 154 Belgian pig holdings in association with myocardial failure and sudden death in fatteners and suckling piglets or with reproductive failure in sows. To clarify some epidemiological aspects 3 EMCV isolates characteristic for the different clinical pictures and outbreaks were studied. Field observations and animal experiments indicated that the pathogenicity induced by each isolate is specific for one age category and that the spread of the virus is limited. The presented data also suggest that rodents may play a role in the transmission of EMCV but that pig-to-pig transmission is at least as important. Molecular analysis of two separate regions on the genomes of the respective EMCV isolates showed that the 1995-96 EMCV epizootic in Belgium was due to a new virus introduction. Furthermore, the VP1 coding gene is proposed as a marker of virulence. 相似文献
15.
猪脑心肌炎病毒抗体间接ELISA方法的建立及应用 总被引：1，自引：0，他引：1
应用纯化的EMCV VP1重组蛋白建立了检测EMCV抗体的间接ELISA方法,并确定了间接ELISA的最佳工作条件。结果表明,重组蛋白抗原的最适包被浓度为0.54μg/mL;与间接免疫荧光试验比较,结果表明该方法具有良好的特异性和敏感性。此方法中VP1抗原最佳稀释度为1:100,待检血清的最佳稀释度为1:50,酶标二抗的最佳稀释度为1:800,1%明胶为封闭液,OD450时阴阳性血清的临界值为0.35。对2006年天津7个地区66个猪场295份屠宰猪的血清样本的检测结果显示,天津各地区血清EMCV抗体阳性率在41.67%～93.33%之间,平均84.75%,猪场抗体阳性率在50%～100%,平均93.94%。 相似文献
16.
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
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Characteristics of 2 encephalomyocarditis virus (EMCV) isolates (MN-25 and MN-30) recovered from aborted swine fetuses were examined along with 2 other swine isolates (NVSL-MDV and NVSL-PR) and a reference ATCC strain (VR-129). All 5 EMCV isolates were found to be serologically related by cross testing, using serum neutralization and fluorescent antibody assays. Hemagglutination (HA) properties of the 5 isolates were compared, using 5 diluents. The MN-25 and NVSL-MDV strains had HA activity with guinea pig RBC in all 5 diluents, whereas MN-30, NVSL-PR, and VR-129 had HA activity only in KCl-borate buffer. The HA ability with RBC of various animal species was examined, using KCl-borate diluent. All virus isolates had high HA titer (1:512 to 1:2,048) with guinea pig, rat, and horse RBC and lower HA titer (1:16 to 1:64) with sheep RBC. The MN-25 and NVSL-MDV isolates agglutinated dogs RBC, whereas MN-30, NVSL-PR, and VR-129 strains did not. Viral replication was evident in 8 of 10 cell lines tested, although infectivity titers of each virus varied by cell line used. Plaque-forming ability was similar for all 5 isolates, but plaque size was different by virus and cell culture used. Virus isolates were found to be stable after being heated at 56 C and subjected to a wide range of pH. A viral polypeptide pattern difference for all 5 isolates was not found by use of sodium dodecyl sulfate-polyacrylamide gel electrophoresis.(ABSTRACT TRUNCATED AT 250 WORDS) 相似文献
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对江西省某家养野猪场临诊疑似副猪嗜血杆菌（Haemophilusparasuis,Hps）感染的病例进行细菌分离鉴定,PCR扩增分离菌株的16SrRNA并进行测序分析,并对分离菌进行细菌形态、生化鉴定和PCR鉴定及序列比对分析。结果显示,获得1株家养野猪源Hps分离株（命名为HPJXYZ01）,该分离株与国内外参考菌株序列之闻的同源性为93．1％～99．2％,与本实验室江西省家猪源分离株的同源性为84％～92．1％。结果表明,江西省家养野猪中存在Hps感染,分离株与国内外家猪源Hps间的16SrRNA序列差异不大,Hps16SrRNA核苷酸序列比较稳定,其进化不存在明显的地域相关性。 相似文献
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试验旨在研究杂交野猪猪繁殖与呼吸综合征病毒(porcine reproductive and respiratory syndrome virus, PRRSV) 辽宁分离株的遗传变异情况及分子生物学特征.用Marc-145细胞从辽宁某杂交野猪场疑似猪繁殖与呼吸综合征(porcine reproductive and respiratory syndrome, PRRS)病猪血液中分离到1株病毒,该分离毒株经Marc-145细胞6次传代后出现稳定的细胞病变,采用RT-PCR方法对分离病毒进行ORF6和ORF7基因的扩增、克隆和测序,并与已知序列毒株的相应片段进行同源性比对.结果表明,分离毒株的ORF6、ORF7基因与国内外美洲型毒株的核苷酸同源性分别为96.0%～100.0%、94.5%～99.4%;氨基酸同源性分别为89.6%～100.0%、87.3%～98.7%;与欧洲型代表毒株LV的ORF6、ORF7基因差异较大,核苷酸同源性分别为70.4%、70.1%,氨基酸同源性分别为48.8%、49.7%.推测辽宁杂交野猪体内分离毒株在基因型上属于美洲型毒株. 相似文献