共查询到20条相似文献，搜索用时 31 毫秒
1.
Genetic diversity of avian infectious bronchitis virus isolates in Korea between 2003 and 2006 总被引：1，自引：0，他引：1
Thirty-three field isolates of avian infectious bronchitis virus (IBV) were recovered from commercial chicken flocks in Korea between 2003 and 2006 and were characterized phylogenetically by nucleotide sequence analysis of the IBV S1 gene hyper-variable region. Our phylogenetic analysis revealed that recent field isolates of IBV formed at least three distinct phylogenetic types, including K-I, K-II, and K-III. K-I type IBV consisted of indigenous, 13 IBV isolates which evolved from the Kr-EJ/95 strain and then separated into the lineages of type K-Ia and type K-Ib. K-II type IBV isolates (n = 19) were closely related to nephropathogenic IBV variants from China and Japan. The K-III type isolate (Kr/D064/05), first identified by this study, was closely related to enteric IBV variants from the Chinese strains that cause proventriculitis. Sequence comparisons showed amino acid differences of >27.5% between IBV types. The molecular epidemiologic characteristics of IBV field isolates are briefly discussed. 相似文献
2.
QU Su-jie SHI Kai-chuang SU Yan-qiong HU Jie MO Sheng-lan ZHANG Bu-xian LI Jun ZOU Lian-bin 《中国畜牧兽医》2015,42(12):3119-3125
To investigate genetic variation of avian infectious bronchitis virus (IBV) in Guangxi province,one strain of IBV was isolated from chicken.Two pairs of primers for amplifying the N and M genes of IBV were designed according to the sequences in GenBank.The N and M genes of the strain were amplified by RT-PCR,and they were proved to be the N and M genes of IBV by cloning,sequencing and compared with reference IBV strains published in GenBank.The results showed that the N gene from the IBV isolate consisted of 1 230 bp,coding 409 amino acids.The M gene from the IBV isolate consisted of 678 bp,coding 225 amino acids.The sequence analysis of N gene showed that it shared 87.2% to 93.3% nucleotide homologies and 90.0% to 94.4% deduced amino acid sequence homologies with IBV strains from GenBank.The M gene sequence analysis showed that it shared 83.6% to 91.0% nucleotide homologies and 82.7% to 92.9% deduced amino acid sequence homologies.The phylogenetic tree analysis showed that it was closely related to BJ and LX4 strains,and were clustered into one group;But with the distant relatives from other strains of IBV.These results suggested that the isolate was a new variant of IBV. 相似文献
3.
鸡传染性支气管炎病毒CQ/01/2004株的分离与鉴定 总被引：2，自引：0，他引：2
2004年从重庆某肉用鸡场疑似鸡传染性支气管炎的病鸡中采集病料，按常规处理后接种9～10日龄鸡胚，通过鸡胚连续传代培养3代，并对该分离毒株的鸡胚致病性、血凝性和NDV的干扰特性进行检测．同时进行了动物回归试验。结果表明，该分离株具有IBV感染特征，可使鸡胚胚体出血、蜷缩、矮化；该分离毒株无直接血凝性，对NDV有明显的干扰作用；动物回归试验中有75％的感染鸡在10d内发病或死亡。剖检病死鸡可见肾苍白、肿胀，肾小管内充塞大量尿酸盐，支气管有出血点、有大量粘液。采用反转录．聚合酶链式反应（RT-PCR）技术对CQ／01／2004的纤突蛋白S1基因进行扩增、克隆和序列测定，结果表明该基因具有IBVSl基因的共有分子特征，将测序结果提交GenBank进行同源性检索，发现分离株CQ／01／2004和J株S1的同源性最高，核苷酸同源性为94％，氨基酸同源性为89．4％，与M41的核苷酸同源性为80．6％，氨基酸同源性为78．0％。试验结果表明，分离的病毒株CQ／01／2004为鸡传染性支气管炎病毒。 相似文献
4.
为调查辽宁地区鸡传染性支气管炎病毒(IBV)的流行及遗传演化情况,从辽宁某鸡场疑似鸡传染性支气管炎病料中分离得到1株病毒,经分子生物学检测、鸡胚矮小化试验、新城疫病毒血凝特性干扰试验和动物回归试验,确定该毒株为IBV,并命名为CH/LN/2019。扩增该毒株S1基因并进行序列分析发现,分离株S1基因全长1 620 nt,编码540个氨基酸,S1蛋白裂解位点为HRRRR,属于基因Ⅰ型(QX型)毒株;同源性比对发现,分离株与基因Ⅰ型代表毒株QXIBV株的核苷酸和氨基酸序列同源性最高,分别为95.6%和94.8%;与国内常规型疫苗H52、H120和Ma5毒株的同源性较低,核苷酸和氨基酸序列同源性仅为77.2%~76.9%和75.4%~76.2%。本试验为辽宁地区鸡传染性支气管炎的流行病学调查及免疫防控提供了参考。 相似文献
5.
为研究鸡传染性支气管炎病毒(IBV)广西流行株的遗传变异情况,本研究从广西发病鸡中分离鉴定了1株IBV。参照GenBank中IBV的核苷酸序列设计2对引物,利用RT-PCR技术对分离毒株的N、M基因进行了克隆、序列测定,并与GenBank中发表的国内外参考毒株进行比对分析。结果显示,N基因序列全长为1 230 bp,编码409个氨基酸,M基因序列全长为678 bp,编码225个氨基酸。与参考毒株相比,分离株的N基因核苷酸序列同源性为87.2%~93.3%,推导的氨基酸序列同源性为90.0%~94.4%;M基因的核苷酸序列同源性为83.6%~91.0%,推导的氨基酸序列同源性为82.7%~92.9%。在遗传进化树中,本试验分离株Guangxi156株与BJ株和LX4株两个参考株位于同一个分支上,亲缘关系较近,而与其他参考株属于不同的分支,亲缘关系较远。结果表明,本试验分离株是一株新的IBV变异株。 相似文献
6.
Isolation and identification of four infectious bronchitis virus strains in China and analyses of their S1 glycoprotein gene 总被引：4，自引：0，他引：4
Between 2003 and 2005, four strains of infectious bronchitis virus (IBV) were isolated from the vaccinated chicken flocks in China. The results from chicken embryo cross-neutralization assays showed that all the four isolates were relative to strain A2 of IBV, which was isolated in 1996 in Beijing and related to strain 4/91. The S1 gene of the spike protein was amplified and sequenced. The nucleotide and amino acid sequence of the S1 gene had a similar degree of identity (88.98-99.28%) among the four Chinese IBV isolates. The identity of the S1 protein gene between the four Chinese IBV isolates and 14 strains of other IBVs varied from 70.06 to 81.59%. Phylogenetic analysis suggested that there are at least four groups of IBVs circulating in China and the disease outbreaks might have been caused by infection of multiple strains of IBV. 相似文献
7.
Infectious bronchitis was diagnosed in 3-to-4-week-old pullets from an outbreak in a commercial flock in California. The disease was characterized by head swelling, watery discharge from the eyes and nostrils, and urates in kidneys. Mortality ranged from 1.8% to 12.5% per week. The isolation of a coronavirus from a suspension of pooled kidneys from clinically ill chickens at the fifth passage in 10-day-old chicken embryos, gross and histologic renal lesions, and seroconversion by enzyme-linked immunosorbent assay in inoculated birds suggested that the virus isolated was a nephrotrophic strain of infectious bronchitis virus (IBV). The virus isolate was found to be a previously unrecognized serotype, based on virus neutralization tests performed in embryonated chicken eggs. Nephropathogenicity of the IBV isolate was confirmed by inoculation of the viral isolate into specific-pathogen-free chicks and demonstration of renal lesions. The isolation of nephrotropic strains of IBV has not been reported previously from poultry in California. 相似文献
8.
本研究对分离自沈阳地区的一株传染性支气管炎病毒（ＳＹ毒株）进行了生物学特性的研究,同时成功地对其免疫原Ｓ１基因进行了ＲＴ－ＰＣＲ扩增、克隆与序列分析。 通过电镜观察、动物回归试验、血凝特性研究等试验验证分离自沈阳地区的ＳＹ毒株确实为一株传染性支气管炎病毒。气管环组织培养交叉中和试验结果表明,分离株ＳＹ株不同于参考毒株澳大利亚Ｔ、Ｈ５２、Ｍ４１,且不同于国内其它流行株ＨＤ、ＨＢ、ＸＢ、ＤＢ等,是一个新的变异株。 利用ＩＢＶ Ｓ１基因特异性寡聚核苷酸引物,经ＲＴ－ＰＣＲ扩增ＳＹ毒株的Ｓ１基因,得到预期的约１．７Ｋｂ片段;并将扩增所得ｃＤＮＡ插入克隆质粒ｐＵＣ１９的ＥｃｏＲⅠ／ＢａｍＨⅠ位点,在大肠杆菌ＤＨ５ａ中实现目的基因的克隆。经限制性核酸内切酶分析及ＰＣＲ鉴定,证实为阳性重组质粒,利用末端双脱氧链终止法对其测序,得到Ｓ１基因全长１６４０ｂｐ,包括整个开放阅读框。通过序列分析软件ＤＮＡＳＩＳ、ＰＲＯＳＩＳ、ＭＥＧＡ等软件对Ｓ１基因核苷酸序列及推导的氨基酸序列进行分析,结果表明：分离株ＳＹ与７株参考株和国内流行株ＨＤ株相比,无论是核苷酸序列同源百分率还是氨基酸序列同源百分离都较低,均未达到８０％,这就提示我们ＳＹ毒 相似文献
9.
腺胃病变型鸡传染性支气管炎病毒的分离鉴定 总被引：36，自引：8，他引：28
从１９９７年在辽宁省大连市某鸡场发生的一种以腺胃肿大为特征的鸡的传染病病例中收集腺胃组织（Ｄ９７１）接种ＳＰＦ鸡胚,传至１３代,具有规律性死亡和典型胚胎病变特征,ＥＩＤ５０为１０－６．４８／０．２ｍｌ。Ｄ９７１毒株感染ＳＰＦ鸡胚尿囊液经负染电镜观察,可见直径为８０～１２０ｎｍ,有囊膜和纤突的冠状病毒粒子。Ｄ９７１毒株经卵磷脂酶Ｃ处理后能凝集鸡红细胞,并能特异的被Ｄ９７１多抗所抑制。用反转录聚合酶链反应获得了Ｄ９７１毒株免原（Ｓ１）基因ｃＤＮＡ,经１％琼脂糖凝胶电泳可见１．７ｋｂ的特征性条带。用Ｄ９７１毒株接种ＳＰＦ鸡,能引起与自然病例相似的病理变化,并分离到病毒。用Ｄ９７１毒株可直接感染ＳＰＦ鸡胚成纤维细胞。结果证实从腺胃病变型ＩＢ鸡群中分离的Ｄ９７１毒株是冠状病毒科ＩＢＶ成员。 相似文献
10.
One nephropathogenic infectious bronchitis virus （IBV） strain was isolated from Qingdao city, named as QD isolate.S1 gene of the strain was amplified, cloned and sequenced. The S1 gene of QD isolate was composed of 1620 nucleotides, and a spike glycoprotein cleavage recognition site was Arg-Arg-Phe-Arg-Arg. The nucleotide acid similarities among the nine IBV vaccine strains and the QD strain were 78.8% to 82.2%. Phylogenetic analysis based on the S1 genes showed that QD strain and the vaccine strains belonged to different clusters, and showed larger evolutionary distances, but showed the smaller evolutionary distances with the field nephropathogenic IBV strains in China. The result showed that nephropathogenic IBV strains were widely popular in China, and the QD strain could be used as the infectious bronchitis vaccine candidate strain. 相似文献
11.
本文以嗜肾型鸡传染性支气管炎病毒(Infectious bronchitis virus,IBV)毒株的RNA为模板,通过RT-PCR扩增获得IBV非结构蛋白5(non-structural protein,nsp5)基因片段后,构建了杆状病毒重组质粒IBVnsp5-Bacmid。IBVnsp5-Bacmid转染Sf9细胞,获得nsp5重组杆状病毒,经(immunofluorescence assay,IFA)和Western blot检测到转染细胞表达的nsp5蛋白。进一步从感染的细胞中纯化重组蛋白,并用纯化的蛋白免疫小鼠制备了抗nsp5的多抗血清,该多抗血清可检测到IBV四川株SC021202感染的DF-1细胞中特异性的nsp5蛋白。结果表明IBV nsp5在Bac-to-Bac真核表达系统中获得了成功表达,而且具有良好的免疫原性和反应原性。 相似文献
12.
OBJECTIVE: To identify and partially characterize a coronaviruslike virus isolated from naturally infected pigeons. ANIMALS: 50 specific pathogen-free (SPF) embryonated chicken eggs, 30 White Leghorn SPF chickens, and 12 clinically normal pigeons. PROCEDURES: Pancreatic tissue specimens from sick pigeons were inoculated into SPF embryonated chicken eggs for viral isolation and investigation of morphologic and hemagglutinating properties of the isolate, called PSH050513. Furthermore, virulence studies in SPF chickens and experimental pigeons were performed. The spike (S) glycoprotein gene of PSH050513 was further sequenced and analyzed. RESULTS: PSH050513 was isolated and identified from the experimentally infected pigeons by a routine method, which was in accordance with Koch's postulates. The complete S protein (1,167 amino acids) was compared with published S protein sequences of other avian and mammalian coronaviruses. A high degree of sequence identity (79.3% to 99.6%) was observed between the S protein sequence of PSH050513 and published sequences of avian infectious bronchitis virus (IBV); only limited identity (< 37.8%) was observed with turkey coronavirus and mammalian coronaviruses. Furthermore, when the virus was inoculated into SPF chickens, pancreatitis developed. CONCLUSIONS AND CLINICAL RELEVANCE: PSH050513 has been tentatively identified as a novel member of group 3 coronaviruses that have close genetic relationships with IBV strains. 相似文献
13.
Fragments within S1 genes ((poly100)S1) of infectious bronchitis virus (IBV) strains ZJ971, M41 and SC021202 (SC) were subcloned into a prokaryotic expression vector and expressed in Escherichia coli. Monoclonal antibodies (mAbs) against the recombinant (poly100)S1 proteins were produced, characterized and used to analyse epitopes on the S1 subunit of IBV. Nine mAbs raising from the three (poly100)S1 proteins recognized five different epitopes of the S1 subunit, designated as S1-A, B, C, D and E. Epitopes S1-C and S1-D are common for the three IBV strains, while S1-A and S1-B exist on ZJ971 and M41 strains, and S1-E was a strain-specific epitope for SC strain. Immunocytochemistry indicated that all the mAbs to the (poly100)S1 proteins can react with the homologous S1 glycoprotein expressed in Vero cells. Moreover neutralization test demonstrated that only mAbs 6E2, 4F9 and 6G4 had neutralization activity for the homologous IBV. These mAbs to (poly100)S1 protein were potential candidates for detecting and distinguishing IBV strains, and also used to examine antigenic variation of the S1 protein. 相似文献
14.
15.
鸡传染性支气管炎病毒HN99株S1基因的克隆与序列测定 总被引：1，自引：0，他引：1
根据基因库中收录的鸡传染性支气管炎病毒 (IBV)S1基因的序列 ,设计了一对引物并采用RT -PCR扩增了鸡传染性支气管炎病毒HN99株的S1基因 ,扩增产物进行了克隆、测序 ,获得了IBVHN99株S1基因片段 ,其大小为1 739bp(含前导序列 ) ,其核苷酸序列与H1 2 0、H52、M41、Gray、Holte的S1基因核苷酸序列同源性较低 ,分别为 79.1 %,79.2 %,77.3%,77.8%,79 .4%,有大量的点突变并伴有基因插入和缺失 ;IBVHN99株的S1基因推导的氨基酸与H1 2 0、H52、M41、Gray、Holte株氨基酸的同源性分别为80 .1 %,79.9%,79.5%,78.5%,78.5%,经S1基因系统进化分析 ,提示IBVHN99株与其它各毒株的亲缘关系较远 ,初步证实IBVHN99株为一新的IBV毒株 相似文献
16.
In order to verify a commonly held assumption that only Massachusetts (Mass) serotype of infectious bronchitis virus (IBV) was prevalent in the United States between the 1930s (when IBV was first isolated) and the 1950s (when the use of commercial IBV vaccines began), we examined 40 IBV field isolates from the 1940s. Thirty-eight of those isolates were recognized as Mass serotype viruses based on their reactivity to Mass-specific monoclonal antibody (Mab) and neutralization by Mass-specific chicken serum. The remaining two isolates, N-M24 and N-M39, that did not react with Mass-specific Mab, resisted neutralization by Mass-specific chicken serum, and were neutralized only by homologous chicken antibody were identified as non-Mass IBV. When the first 900 nucleotides (nt) from the 5'-end of the spike (S1) glycoprotein gene and their deduced amino acid (aa) sequences were compared, the two non-Mass isolates differed from each other by 24% and 28%, respectively. In a similar comparison, the non-Mass viruses N-M24 and N-M39 differed from M28, a Mass-type isolate from the 1940s, by 21% and 22% (nt) and 28% and 27% (aa), respectively. These data indicate that antigenic and genetic diversity among IBV isolates existed even in the 1940s. Interestingly, when the N-terminal region of the S1 of M28 was compared to that of M41, a prototype Mass virus that has undergone countless number of in vivo and in vitro host passages, the two viruses differed by only 2% (nt) and 4% (aa). This finding suggests that frequent genetic changes are not inherent in all IBV genomes. 相似文献
17.
根据已发表的鸡传染性支气管炎病毒S1基因,设计并合成了一对引物,经RT-PCR扩增后获得全长约为1700bp的核苷酸序列。序列分析表明,S1基因的最大开放阅读框位于12位～1685位碱基之间,编码557个氨基酸;KIBVXJ株S1基因序列在19位～292位点的氨基酸区域内有较多的氨基酸置换、插入和与缺失现象;S1的裂解位点的序列为HRRRR,具有我国地方流行株的特征。KIBVXJ株的S1基因与国内外疫苗株的同源性分析表明,核苷酸同源率为73.8%～74.2%,氨基酸同源率为74.9%～76.0%。遗传进化分析表明,KIBVXJ株与国内外的主要疫苗株亲缘关系最远,与国内近年来分离的A2株、LX4株和QXIBV株的亲缘关系最近。 相似文献
18.
Between 2006 and 2009, seven strains of infectious bronchitis (IB) virus (IBV) were isolated from vaccinated chicken flocks on different chicken farms in China. The pathogenic characters of seven IBV strains were assessed. Each of the seven strains was infective to the test chickens and could induce an immune response. The results from chicken embryo cross-neutralization assays showed that these strains were antigenically distinct from classic IBV strains of H120, M41, Conn, and Gray. Compared to H120 vaccine strain, point mutation, short insertion, and deletion occurred at many positions in the S1 protein of the seven strains. Five of the seven strains had the motif (HRRRR), which was identical to that of the epidemic IBV strains in China. Two new motifs (HRLRR and RRIRR) emerged in the isolated strains. The homology of the nucleotide and amino acid sequences of the S1 gene among the seven isolates was 81.7%-99.7% and 79.0%-99.4%, respectively. These seven strains were also genetically different from the vaccine strains and non-China IBV strains but closely related to large numbers of Chinese strains. The seven isolates and 36 reference IBV strains were clustered into six distinct groups (I-VI). The seven strains were categorized into groups I, II, and III, forming a big phylogenetic branch, which is closely related to Chinese IBVs, whereas the vaccine strains belonging to group VI are genetically distant from groups I, II, and III. The results from this study indicate that different IBV strains cocirculate in the chicken population in China. 相似文献
19.
从黑龙江省绥化市某养鸡场疑似鸡传染性支气管炎的鸡群中采集病料,经鸡胚传代、电镜观察、生物学特性和分子生物学鉴定以及动物回归试验,表明该分离株具有明显的鸡传染性支气管炎病毒特征,具有鸡胚出现卷曲、出血、发育延迟等作用;对新城疫病毒有明显的干扰作用;动物回归试验导致未免疫鸡出现明显的感染症状,剖检可见明显的肾脏肿胀、尿酸盐沉积现象,同时可见呼吸道有出血现象。对该毒株的N基因进行扩增,并将其序列与NCBI的参考毒株的N基因进行序列对比,发现其与国内的分离株SC和N毒株的N基因具有较高的同源性,为97.6%;与国外参考毒株和国内的疫苗株同源性较低,为84.8%。表明分离的病毒为鸡传染性支气管炎病毒。 相似文献
20.
鸡传染性支气管炎病毒SC株N基因序列测定与同源性分析 总被引：1，自引：0，他引：1
从我国四川省一疑似鸡传染性支气管炎(Avianinfectious bronchitisvirus,IB)的雏鸡病料中成功分离到一株鸡传染性支气管炎病毒(Infectious Bronchitis Virus,IBV),命名为SC。病毒经鸡胚传代、血凝试验监测和负染电镜检查证实为IBV。自接毒鸡胚尿囊液中提取RNA后应用反转录-聚合酶链反应(RT-PCR)扩增得到了IBVSC株mRNA6 cDNA(编码N蛋白)。应用DNAstar5．06,Clustal1．8分析软件将克隆测序的N蛋白基因与Genbank中11株国内外参考毒株进行序列比较分析和同源性分析,发现IBV SC株变异独特,明显不同于国内外参考毒株。 相似文献