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1.
CIAV-7 is a virus with similar pathogenic and physicochemical characteristics to, but antigenically distinct from, chicken infectious anemia virus (CIAV). The pathogenesis of CIAV-7 was evaluated in a comparative study with a representative isolate of CIAV, the Del-Ros strain. The pathogenesis of CIAV-7 was similar to Del-Ros on the basis of the clinical disease induced and gross and microscopic lesions, although CIAV-7 produced fewer and less severe lesions overall. A second comparative pathogenesis study was performed with Del-Ros and CIAV-7, both alone and in combination with infectious bursal disease virus (IBDV). In this study, the pathogenesis of CIAV-7 was similar to Del-Ros in clinical, gross, and microscopic lesions in the bone marrow. However, thymic lesions were less severe in CIAV-7-inoculated birds. The interaction between Del-Ros and IBDV was synergistic, whereas there was no observed potentiation of CIAV-7-induced disease by IBDV. Progeny from breeder flocks from several geographic locations in the eastern United States were challenged with CIAV-7 or Del-Ros to assess protection by maternal antibodies. Some progeny from all flocks had protection against CIAV-7 challenge, providing evidence for the presence of CIAV-7 in the field. Additionally, the number of birds protected against CIAV-7 or Del-Ros challenge varied within flocks, demonstrating that the agents are serologically distinct. 相似文献
2.
Seven chicken infectious anemia virus (CIAV) isolates were obtained from seven broiler flocks with poor performance in two states of Brazil. All isolates induced thymus atrophy, bone-marrow aplasia, and low hematocrit values when inoculated into 1-day-old susceptible chicks. The CIAV isolates were resistant to treatment with chloroform and were able to pass through 50-nm-pore-size filters. CIAV-specific antigens could be demonstrated in tissues of experimentally infected chicks using a monoclonal antibody specific for CIAV. These characteristics of the virus and the virus-induced lesions demonstrate that CIAV is present in Brazil and that the virus is associated with production problems. 相似文献
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采用PCR方法，以传染性贫血病毒(CIAV)DNA为模板，扩增并克隆了CIAV的VP1、VP2基因，并进行了序列分析。经基因修饰，将这两个基因分别加上表达元件后连接作为目的基因。通过同源重组技术，将目的基因插入到火鸡疱疹病毒(HVT)gC基因区，构建了一株含CIAV VP1、VP2基因表达单元的重组HVT(VP1VP2-rHVT)：体内、体外传代结果表明该重组病毒性状稳定。采用PCR扩增及Southem blot杂交检测，证实了CIAV VP1、VP2基因的插入。 相似文献
4.
本试验根据GenBank中登录的禽传染性贫血病毒(CAV)基因序列,设计合成2对引物,外引物的扩增片段大小为485 bp,内引物的扩增片段大小为297 bp,建立了适合CAV快速检测的套式PCR方法(nested PCR),并采用该方法对CAV阳性毒株及临床病料进行了检测。结果显示,该方法能扩增到297 bp的条带,禽流感病毒(H9亚型)、新城疫病毒、传染性法氏囊病病毒、禽网状内皮增生病病毒、减蛋综合征病毒、禽呼肠孤病毒、马立克氏病病毒的扩增结果均为阴性。该方法第1步扩增的敏感性是100 pg,第2步PCR扩增的敏感性是1 fg,敏感性提高了105倍。本研究建立的CAV套式PCR方法具有敏感性高、重复性好、特异性强等优点,可用于CAV的临床诊断和分子流行病学调查等。 相似文献
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Coinfection of specific-pathogen-free chickens with Marek's disease virus (MDV) and chicken infectious anemia virus: effect of MDV pathotype 总被引：3，自引：0，他引：3
Both Marek's disease virus (MDV) and chicken infectious anemia virus (CIAV) infections are prevalent in chickens throughout the world. In the past decade, MDV strains with increased virulence (very virulent plus MDV pathotype [vv+MDV]) have been isolated. The purpose of this experiment was to determine the effects of coinfection of chickens with CIAV and a vv+MDV isolate. Specific-pathogen-free chickens were inoculated at 1 day posthatch with RB1B (very virulent MDV pathotype [vvMDV]) only, 584A (vv+MDV) only, CIAV only, RB1B + CIAV, 584A + CIAV, or nothing. Samples of spleen, thymus, and bursa of Fabricius were collected at 4, 7, 10, and 13 days postinoculation (DPI). Thymic and bursal atrophy at 13 DPI and final mortality at 30 DPI were significantly greater in chickens inoculated with 584A with or without added CIAV, or with RB1B plus CIAV, compared with birds inoculated with RB1B alone. Both amounts of virus reisolated and levels of virus detected by quantitative-competitive polymerase chain reaction were greater at 4 DPI in 584A inoculates compared with RB1B inoculates. To monitor the early cytolytic infection, northern analysis was done with a probe for the MDV immediate early gene ICP4 (infected cell protein 4). In the absence of CIAV, ICP4 expression was more apparent in chickens inoculated with 584A than in those inoculated with RB1B. CIAV coinfection increased ICP4 expression in the spleens of chickens infected with RB1B. These results indicated that inoculation of chickens with the 584A isolate caused a more robust early cytolytic infection compared with inoculation with RB1B alone and support the classification of 584A as a vv+MDV strain. Coinfection with CIAV exacerbated vvMDV strain RB1B infection. The extent of this exacerbation was less evident when birds were coinfected with 584A and CIAV. 相似文献
7.
鸡传染性贫血病毒与禽网状内皮增生症病毒二重PCR检测方法的建立 总被引：3，自引：3，他引：0
当前养鸡业中鸡传染性贫血病毒、禽网状内皮增生症病毒混合感染的发生率日益增高,为提高CIAV、REV检测效率,根据GenBank上登录的CIAV全基因序列和REV的LTR序列分别设计了2对引物,建立了REV、CIAV二重PCR检测方法,另外还比较了2种不同的DNA提取方法。结果显示,该方法扩增目的条带清晰、敏感性高、特异性好。应用建立的方法对临床7份病料样品检测,检出5份CIAV、REV阳性,表明建立的二重PCR方法能有效用于CIAV、REV混合感染的临床诊断。 相似文献
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Recombinant Protein A, recombinant Protein G, and anti-chicken-IgG anti-bodies raised in rabbits, goats, or horses were found to bind directly to chicken infectious anemia virus (CIAV). MSB-1 cells infected with the Cux-1 strain of chicken anemia agent, but not to uninfected MSB-1 cells were found to react with fluorescein isothiocyanate conjugates. In an enzyme-linked immunosorbent assay, rabbit anti-chicken horseradish peroxidase conjugate bound directly to CIA-1 CIAV-coated plates. In addition, sera from a low percentage of specific-pathogen-free breeder hens reacted in an indirect fluorescent antibody test to detect CIAV antibodies. These reactions generally disappeared within a month. The breeder flocks were demonstrated to be free of CIAV infection by the susceptibility of their progeny. 相似文献
11.
Chicken infectious anemia virus (CIAV) is a ubiquitous and highly resistant virus of chickens that causes anemia and death in chicks less than 3 wk of age and immunosuppression in chickens older than 3 wk of age. The production of specific-pathogen-free eggs free of CIAV is essential for research and vaccine production. Currently, flocks are screened for CIAV by antibody tests to ensure freedom from CIAV infection. Recent evidence, however, indicates that chickens may carry and vertically transmit CIAV DNA independently of their antibody status. In this study, we tested embryos and eggshell membrane residues by nested polymerase chain reaction (PCR) as a sensitive method of detecting CIAV DNA. CIAV DNA could be detected in the blastodisks and semen obtained from antibody-positive and -negative chickens. Examination of different tissues between 18 and 20 days of incubation indicated that many but not all organs of individual embryos were positive. The lymphoid organs and gonads had the highest incidence of CIAV DNA, which was significantly different (P < 0.05) from the incidence in the liver. Eggshell membrane samples from embryos or newly hatched chicks were an excellent noninvasive source for the detection of CIAV DNA, identifying significantly more positive embryos than did pooled lymphoid organs. The use of dexamethasone injections as a method to improve the detection of carrier birds did not result in an increase of vertical transmission or cause seroconversion in the treated hens. A combination of testing eggshell membrane residues at hatch and periodic testing of blood DNA by nested PCR can be used to identify chickens carrying CIAV DNA and may be used to eradicate carrier birds. 相似文献
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The Becht strain of infectious bursal disease virus was compared with a virus isolated from Aedes vexans mosquitoes and designed 743 virus. The viruses were compared with respect to cell culture host range, cellular changes resulting from viral infections, growth curves, antigenic relationship, and physicochemical characteristics. The viruses are closely comparable in all these properties, and they are considered to be strains of the same virus. In cross comparisons by the enzyme-linked immunosorbent assay, 743 virus and infectious bursal disease virus were found to be antigenically identical, confirming the results of the neutralization test. The 743 virus differs from most strains of infectious bursal disease virus in that it is nonpathogenic for chickens. 相似文献
13.
Establishment of conditions for the detection of bovine herpesvirus-1 by polymerase chain reaction using primers in the thymidine kinase region. 
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下载免费PDF全文 C V Yason L M Harris P K McKenna D Wadowska F S Kibenge 《Canadian journal of veterinary research》1995,59(2):94-101
Polymerase chain reaction (PCR) for detection of bovine herpesvirus-1 (BHV-1) was developed and optimized using 22 bp sense and 20 bp antisense primers in the thymidine kinase (TK) coding region. The amplification product is 183 bp long. The PCR optimization was done using BHV-1 tissue culture supernate (BHV-1TCS), concentrated BHV-1 tissue culture supernate (cBHV-1TCS) and sucrose gradient purified BHV-1 (pBHV-1). The sensitivity of four methods of sample preparation which are standard DNA extraction, modified proteinase K (PK) digestion, GeneReleaserTM + 34 cycles or + 44 cycles, and boiling were compared with virus isolation (VI) using BHV-1TCS. The incorporation of 10% glycerol in the reaction mixture, the incubation in PK for 18 hours and predenaturation of samples and cooling in ice prior to PCR were essential for the amplification of BHV-1 DNA for samples prepared by standard DNA extraction and modified PK digestion. The preparation of samples by Gene-ReleaserTM, a proprietary nucleic acid releasing cocktail, showed 10 to 1,000-fold increase in sensitivity compared to standard DNA extraction and modified PK digestion. No amplification was observed in samples prepared by boiling. The sample preparation of BHV-1 LA strain by GeneReleaserTM showed sensitivity equivalent to virus isolation. The BHV-1 TK PCR using GeneReleaserTM has a detection limit of 1 picogram and 10 fentograms of purified BHV-1 DNA using ethidium bromide stained gel and Southern blot hybridization, respectively. It could detect viral DNA in 1,000 infected cells in a total suspension of 10,000 cells using either ethidium bromide stained gel or Southern blot hybridization. 相似文献
14.
Owoade AA Oluwayelu DO Fagbohun OA Ammerlaan W Mulders MN Muller CP 《Avian diseases》2004,48(1):202-205
Sera samples from seven poultry farms in southwest Nigeria consisting of 7 broiler, 10 pullet, 1 layer, 1 cockerel, and 1 broiler breeder flocks were tested for the presence of chicken infectious anemia virus (CIAV) antibodies using a commercial enzyme-linked immunosorbent assay kit. Eleven of the 20 flocks (55%) and six out of seven (86%) farms were positive for CIAV antibodies. The seroprevalence largely depended on the age of the flocks. Seroprevalence was higher within the older pullet and layer flocks (83%-100%) than in the younger broiler flocks (0%-83%). In essence, all flocks older than 6 to 8 wk became infected. This is the first report of serologic evidence of CIAV in Subsaharan Africa. Since Southwest Nigeria is the main port of entry of imported chicken and the hub of major poultry breeders, the disease can probably be found throughout the country and beyond. Further studies are necessary to assess economic losses due to CIAV and the cost benefit of countermeasures. 相似文献
15.
Comparative susceptibility of Marek's disease cell lines to chicken infectious anemia virus 总被引：4，自引：0，他引：4
Calnek BW Lucio-Martinez B Cardona C Harris RW Schat KA Buscaglia C 《Avian diseases》2000,44(1):114-124
Chicken infectious anemia virus (CIAV) is known to infect and replicate in various Marek's disease chicken cell lines (MDCCs) derived from Marek's disease (MD) tumors. One line, MDCC-MSB1, has been the substrate used in most studies. We compared a total of 26 MDCCs, including two sublines of MDCC-MSB1, MSB1 (L) and MSB1 (S), four other MD tumor-derived lines, and 20 lines derived from MD virus-induced local lesions, for susceptibility to the Cux-1 and CIA-1 strains of CIAV. The cell lines represented six phenotypic groups of T cells based on the expression of CD4, CD8, and TCR-2 and -3 surface markers. Susceptibility was measured by the number of cells positive for viral antigen in immunofluorescence (IF) tests at 3-10 days postinfection. No clear-cut differences were found in susceptibility related to phenotype, although CD4-/8+ lines and CD4-/8- lines might be more susceptible than CD4+/8- lines. However, several individual lines were more susceptible to Cux-1 than the two MSB1 sublines tested. Contrary to an earlier report, cells of MDCC-CU147, a CD8+, TCR3+, local-lesion derived line, were found to be susceptible to CIA-1. In fact, CU147 was distinguished by very high susceptibility to both CIAV strains. In direct comparisons with MSB1, CU147 detected approximately 10-fold lower doses of virus. Also, virus spread was faster (P < 0.05) in CU147 than in MSB1 and other lines. Results from polymerase chain reaction (PCR) tests to detect infection in titrations were in general agreement with IF test results although PCR detected infection in a few terminal dilution cultures that were negative by IF. 相似文献
16.
Development of a semi-nested PCR for the improved detection of Mycoplasma bovis from bovine milk and mucosal samples 总被引：3，自引：0，他引：3
A new forward primer, Mb-F, was designed to improve the sensitivity and reproducibility of the Mycoplasma bovis-specific PCR developed by Ghadersohi et al. [Vet. Microbiol. 56 (1997) 87] for testing clinical samples. A semi-nested (SN) PCR configuration was developed and this provided enhanced sensitivity and reproducibility. The detection limit of the SN PCR was in the range of 10-100cfu/ml and the correct amplicon was amplified from 9.15pg/microliter of total extracted DNA (mixture of M. bovis and bovine cellular DNA). A dot blot assay was also developed and compared with the SN PCR on a number of randomly selected milk and mucosal samples. The dot blot had the same level of detection as the SN PCR. The specificity of the SN configuration was confirmed by Southern blot analysis and automated sequencing of the PCR product. The results from the tests on the samples from cattle, together with those from sheep, provided evidence that M. bovis is host-specific and that most cattle are colonised. The assay was shown to be specific, sensitive and reproducible and could be used successfully to detect M. bovis directly from clinical material without pre-enrichment. 相似文献
17.
通过对3株鸡传染性贫血病毒(CIAV)的全基因组序列比较分析,部分表明广西南宁鸡群CIAV毒株的遗传变异特征。将阳性CIAV的组织样品进行DNA抽提后,运用PCR方法分段扩增CIAV的全基因组,将获得的PCR产物进行基因克隆并进行阳性克隆菌鉴定后送测序。将所获得的基因序列进行拼接成CIAV基因组全长后,应用LaserGene7.1和MEGA4.1软件对CIAV全基因、Viral protein 3(VP3)、Viral protein 1(VP1)和Viral protein 2(VP2)基因序列进行核苷酸、氨基酸同源性分析和遗传进化分析;并对VP1、VP2和VP3蛋白上与毒力、蛋白磷酸酶活性和细胞凋亡相关的氨基酸位点进行分析。结果获得3株CIAV的全基因序列,分别命名为GX1801、GX1804和GX1810;CIAV全基因遗传进化分析表明GX1801、GX1804和GX1810均同属于Group A群,与国内强毒株GD-103和GD-104毒株的亲缘关系较近;基于VP1基因构建的遗传进化树与全基因构建的进化树最相似;GX1801、GX1804和GX1810 VP1蛋白上与毒力相关位点的第75、89、125、141、144、394位氨基酸位点与日本强毒株C368株、国内强毒株GD-103和GD-104株在同一位点保持一致;GX1801、GX1804和GX1810 VP2蛋白上与磷酸酶活性相关的基序(I^94CNCGQFRKH^103)均未发生变异;GX1801、GX1804和GX1810 VP3蛋白与诱导细胞凋亡相关的重要区域(VP3蛋白N端结构域的第1~69位氨基酸)均高度保守。本研究对3株CIAV全基因组进行测定并进行基因分析,获得了其基因组特征,为进一步研究广西CIAV的分子流行和致病性提供参考。 相似文献
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Characterization of proteins of chicken infectious anemia virus with monoclonal antibodies. 总被引：4，自引：0，他引：4
Eight monoclonal antibodies (MAbs) against chicken infectious anemia virus (CIAV) were developed. These MAbs identified three isolates adapted to grow in the Marek's disease chicken cell line MSB1 (Cux-1, GA-1, and Conn-B) and the chicken-propagated CIA-1 isolate. All MAbs stained MSB1 in the same way with mostly perinuclear staining, although larger nuclear inclusions and cytoplasmic staining were also detected. None of the MAbs neutralized Cux-1. All MAbs reacted in a direct enzyme-linked immunosorbent assay with Cux-1 antigen treated with 0.5% sodium dodecyl sulfate followed by extraction with chloroform, but not with MSB1 cells infected with Cux-1 or chloroform-extracts of these cells. Three viral proteins--VP1, VP2, and VP3--with estimated sizes of 45, 30, and 16 kilodaltons (kd), respectively, were immunoprecipitated using the MAbs and Cux-1-infected cell lysates. The 16-kd protein was the major VP. In addition, a 79-kd protein was detected in infected cell lysates by immunoprecipitation with CIAV-antibody-positive and -negative chicken serum, and CIAV-specific and non-specific MAbs. 相似文献
19.
应用免疫SPA菌体花环法、MTT法和间接ELISA法对鸡传染性贫血病(CIA)-传染性法氏囊病(IBD)联合免疫种鸡后,其子代雏鸡外周血液T细胞数量和增殖功能以及免疫球蛋白IgG、IgM、IgA含量变化进行了动态研究。结果发现,CIA-IBD联合免疫种鸡后,其子代雏鸡外周血液T细胞数量和IgG、IgM、IgA含量均不同程度地高于未免疫的相应对照雏鸡,表明CIA-IBD联合免疫种鸡后,其子代雏鸡外周血液体液免疫和细胞免疫功能明显增强。而CIAV、IBDV强毒攻击后,未免疫的子代雏鸡,其外周血液的上述指标均明显低于疫苗免疫的子代雏鸡,表明未免疫的子代雏鸡外周血液的免疫机能降低,这与未免疫雏鸡缺乏特异性抗体,强毒攻击后,雏鸡免疫器官组织广泛损害,淋巴细胞变性坏死等有关。 相似文献
20.
Davidson I Kedem M Borochovitz H Kass N Ayali G Hamzani E Perelman B Smith B Perk S 《Avian diseases》2004,48(1):108-118
The impact of chicken infectious anemia virus (CIAV) infection on commercial chicken flocks in Israel was examined by analyzing flocks with or without typical CIAV signs, signs of other diseases, or apparently healthy flocks. In 23 flocks (broilers and layers) of ages up to 8 wk, typical signs of CIAV infection (stunting, gangrenous dermatitis, and secondary bacterial infections) were recorded. When permitted by flock owners, in several cases among these 23 flocks the morbidity, mortality, and performance parameters were recorded; the presence of CIAV was detected by polymerase chain reaction (PCR); and the antibody status of parents and broilers was measured. In addition, total mortality, number of birds sold, total kilograms of meat sold, density (kg/m2), mean age at slaughter, daily growth rate in grams, total kilogram of food consumed, food conversion rate, and the European Index were calculated. We also surveyed flocks affected by other diseases, such as tumors, respiratory diseases, or coccidiosis, and flocks with no apparent clinical signs. The latter flocks were negative by CIAV-PCR, indicating that typical CIAV clinical signs are associated with one-step PCR-CIAV amplification. However, a small amount of CIAV might still be present in these flocks, acting to induce the subclinical effects of CIAV infection. These data indicate a link between the presence of virus sequences and typical CIAV signs and strengthen the concept that CIAV infection has a negative economic impact on the chicken industry. 相似文献