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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
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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
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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
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Three coronaviruses isolated from the intestines of laying chickens were partially characterized. Serological and molecular assays indicated that the enteric coronaviruses are infectious bronchitis virus (IBV) isolates. Although the three isolates were recovered from three unrelated chicken flocks, their RNase T1-resistant oligonucleotide fingerprints were almost identical. The three isolates were not neutralized by antisera specific to IBV serotype Connecticut, but their RNase T1-resistant oligonucleotide fingerprints closely matched the fingerprints of strain Conn-46, a Connecticut serotype. This and the co-fingerprinting data suggested that the three field isolates may have emerged from the Connecticut virus through mutation(s). The mutation(s) apparently involved the S1 protein gene that determines the virus serotype. 相似文献
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鸡传染性支气管炎病毒地方流行株的分离与鉴定 总被引：3，自引：0，他引：3
从山西各地区疑似鸡传染性支气管炎(IB)的病料中,分离到5株鸡传染性支气管炎病毒(IBV)分离株,并对分离病毒进行了病毒形态观察、对鸡新城疫病毒(NDV)的干扰、鸡胚致病性试验、动物回归试验、血凝特性试验、病毒理化特性测定等生物特性鉴定及IBV N基因特异性片段的检测.电镜观察,可见直径为60～120 am,有囊膜及纤突呈冠状排列的病毒粒子;对NDV有明显的干扰作用;分离株的传代物均有明显的致鸡胚矮小化作用;动物回归感染死亡鸡肾脏病变明显,表现肾脏肿大、花斑肾现象,输尿管内充塞大量尿酸盐;无直接血凝性,经1%胰酶处理后可凝集鸡红细胞;分离株对乙醚和氯仿敏感;采用反转录-聚合酶链式反应(RT-PCR)对分离毒株进行扩增,结果均扩增出特异N基因核酸片段. 相似文献
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Helena Grgi? D. Bruce Hunter Peter Hunton éva Nagy 《Canadian journal of veterinary research》2009,73(3):212-216
Infectious bronchitis (IB) is an economically important viral disease with worldwide distribution. Every country with an intensive poultry industry has infectious bronchitis virus (IBV). The virus rapidly spreads from bird to bird through horizontal transmission by aerosol or ingestion. Sentinel bird studies were carried out in southern Ontario and IBV has been isolated from layer flocks. Genetic analysis of the S1 region of the strains showed that they were not vaccine related. The pathogenicity of selected Ontario variants of IBV isolates was studied and the subsequent work was to determine the degree of protection against field isolates provided by a commonly used vaccine MILDVAC-Ma5 in Ontario. The protection was evaluated by challenging immunized chickens with the respiratory (IBV-ON1) and nephropathogenic (IBV-ON4) viruses. The mean vaccine efficacy for IBV-ON1 was 66.7% indicating that a Massachusetts serotype vaccine would provide some protection against IBV field isolates. 相似文献
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Takase K Murakawa Y Ariyoshi R Eriguchi S Sugimura T Fujikawa H 《The Journal of veterinary medical science / the Japanese Society of Veterinary Science》2000,62(12):1327-1329
To monitor the existence of avian pathogens in laying chicken flocks, specific pathogen-free (SPF) chickens were introduced into two layer farms and reared with laying hens for 12 months. SPF chickens were bled several times after their introduction and examined for their sero-conversion to avian pathogens. As a result, antibodies to eight or ten kinds of pathogens were detected in SPF chickens on each farm. Antibodies to infectious bronchitis virus (IBV), avian nephritis virus, Mycoplasma gallisepticum and M. synoviae were detected early within the first month. Antibody titer to IBV suggested that the laying chickens were infected with IBV repeatedly during the experiment on both farms. However, antibodies to infectious bursal disease virus and 6 pathogens were not detected. 相似文献
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鸡传染性支气管炎病毒CK/CH/LHLJ/04V的分离鉴定及致弱的初步研究 总被引：1，自引：1，他引：1
从黑龙江省某鸡场发病鸡群的肾脏中分离到一株病毒,通过病毒致SPF鸡胚病变特征、对鸡外周血红细胞凝集特性、病毒粒子形态学特征以及RT-PCR鉴定等方面的研究,表明该病毒为鸡传染性支气管炎病毒,并命名为CK/CH/LHLJ/04V。通过对该病毒基因型分析以及对SPF鸡致病性试验发现该病毒为我国近年来流行的一类重要IBV的代表。将该毒株在SPF鸡胚连续传代110代(P110),取不同代次毒进行动物实验。结果显示,该毒株对SPF雏鸡的致病力随鸡胚传代次数的增加而逐渐降低。P110代毒以105.0 EID50/0.1 mL的剂量通过点眼滴鼻接种15日龄SPF雏鸡,鸡群发病率和死亡率均为0。不同代次毒接种SPF鸡对同源毒株P3代强毒的攻击均具有100%保护性。实验表明,IBV毒株CK/CH/LHLJ/04V P110对SPF雏鸡已无致病性,但仍具有良好的免疫原性,可作为研制IB弱毒疫苗的候选毒株。 相似文献
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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
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巢式PCR快速鉴定鸡传染性支气管炎病毒的研究 总被引：4，自引：0，他引：4
鸡传染性支气管炎病毒（ＩＢＶ）经鸡胚增殖后,直接用尿囊液提取ＲＮＡ后反转录成ｃＤＮＡ,用ＩＢＶ基因３’端的ＵＴＲ１－／ＵＴＲ２＋和ＵＴＲ３－／ＵＴＲ４＋两对引物进行巢式ＰＣＲ,所检测的４个ＩＢＶ标准参考株和１６个ＩＢＶ野毒株均得到了预期的１７４ｂｐ大小的片段,而鸡新城疫病毒（ＮＤＶ）,鸡传染性法氏囊病毒（ＩＢＤＶ）及正常鸡胚尿囊液经同样处理没有可见片段出现。本试验不需纯化册毒只需０５ｍｌ病毒尿囊液即可在２４小时内得到准确的试验结果。这表明与其它ＩＢＶ鉴定方法相比,该法具有快速、灵敏、特异的优点。 相似文献
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Fifty-six isolates of avian infectious bronchitis virus (IBV) were obtained from different field outbreaks in China in 2010, and they were genotyped by comparison with 19 reference strains in the present study. The results showed that LX4-type isolates are still the predominant IBVs circulating in chicken flocks in China, and these isolates could be grouped further into two clusters. Viruses in each cluster had favored amino acid residues at different positions in the S1 subunit of the spike protein. In addition, a recombination event was observed to have occurred between LX4- and tl/CH/LDT3/03I-type strains, which contributed to the emergence of a new strain. The most important finding of the study is the isolation and identification of Taiwan II-type (TW II-type) strains of IBV in mainland China in recent years. The genome of TW II-type IBV strains isolated in mainland China has experienced mutations and deletions, as demonstrated by comparison of the entire genome sequence with those of IBV strains isolated in Taiwan. Pathogenicity testing and sequence analysis of the 3' terminal untranslated region revealed that TW II-type IBV strains isolated in mainland China have a close relationship with the embryo-passaged, attenuated TW2296/95. 相似文献
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Molecular epidemiology of infectious bronchitis virus isolates from China and Southeast Asia 总被引：14，自引：0，他引：14
In order to trace the origin and evolution of avian infectious bronchitis virus (IBV) isolates in China and Southeast Asia, genomic sequencing was used for molecular characterization of 24 IBV isolates and two reference strains in comparison with the published sequences. The 5' region of the S1 genes, containing hypervariable regions I and II, and 3' region of the nucleocapsid genes, containing cytotoxic T lymphocyte epitopes, were used to construct phylogenetic trees for analysis. The results showed that the 24 isolates could be divided into three distinct groups, that is, American, Asian, and European. Some isolates formed a distinct Asian phylogenetic group, suggesting that IBV has existed for some time in Asia. Our results also showed that in vivo recombination of IBV may have occurred at a rather high frequency, contributing to the diversity of these IBV isolates. Importantly, recombination events have probably occurred between vaccine strains and field strains in the natural condition. 相似文献
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对自1994—2009年从我国5省区免疫鸡群中分离到的37株传染性支气管炎病毒(IBV)的S蛋白基因序列进行分析,发现S1基因序列存在广泛的氨基酸替换、缺失和插入现象,大部分IBV分离株S1基因的推导氨基酸序列变异主要集中在60～63、73～74、97、128、282～299位等。S2基因较为保守,主要在裂解位点后的2～47、122～152位发生氨基酸的替换,可见IBV S基因的不断变异可能是造成本试验所调查的5省区免疫鸡群IB频发的重要原因。遗传进化分析发现本试验所调查地区近十多年来肾型毒株仍是主要流行株,没有或少有4/91型毒株流行。所调研地区鸡的腺胃炎持续广泛地发生,但是从临床腺胃病料中很少分离到IBV,可见IBV不太可能是引起腺胃炎的主要病原。 相似文献
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G Liu Q Wang N Liu Y Xiao T Tong S Liu D Wu 《Veterinary immunology and immunopathology》2012,148(3-4):353-358
Infectious bronchitis (IB) is an acute and highly contagious viral respiratory disease of chickens. To understand the kinetics and relationships between the humoral (Ab) and antigen specific T cell immunity as well as pathological changes during infectious bronchitis virus (IBV) infection and immunization, one-week-old SPF chickens were vaccinated with live IBV H52 strain and challenged with IBV M41 15 days post primary infection. Chickens were sacrificed every 3 days to monitor antigen specific serum IgG and IBV nucleoprotein-specific immune responses using a chicken MHC I tetramer developed in our laboratory. The results demonstrated that T cell responses developed more rapidly than the humoral (Ab) immune response after vaccination with H52. However, serum IgG dramatically increased after M41 challenge. Chickens from the control, non-vaccinated group developed severe respiratory symptoms and demonstrated significant pathological changes in lung, kidney and bursa of Fabricius post challenge with M41. However, chickens vaccinated with H52 did not demonstrate clinical signs or histological changes post challenge with M41. These results indicated that the live IBV H52 inoculation effectively protected chickens from morbidity and pathological changes associated with IBV infection. These data facilitates the design of a new generation of IBV vaccine. 相似文献