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From fresh faeces of a wild bird (Melanitta fusca), a virus that showed granular cytopathic effects (CPE) on chicken kidney cell (CKC) cultures was isolated. By indirect immunofluorescence analyses (IFA), this isolate reacted with an antiserum against a bovine rotavirus. The isolate produced clear plaques on CKC by conventional techniques, without trypsin. Three virus plaques were selected by plaque size (small, medium, and large) and cloned by three successive plaque cloning. In the SDS-PAGE analyses, dsRNA bands showed a typical profile of avian rotavirus and quite different from that of avian reovirus. With dsRNA patterns, IFA results, CPE, and a morphological property, the clones were identified as avian rotaviruses of group A rotavirus. The clones killed chicken embryos, when they were inoculated to yolk sac.  相似文献   

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A molecular study of intestinal samples from 21 broiler flocks with a history of enteritis revealed that 23.8% and 14.3% were positive for chicken astrovirus (CAstV) and avian rotavirus (ARV), respectively. CAstV and group A ARV were simultaneously detected in only one broiler flock. Birds in this group developed the significant intestinal lesions characterized by frothy contents, paleness, and thin intestinal walls. In this report we present an unusual case of runting stunting syndrome (RSS) with a history of high mortality and growth retardation in broiler chickens. We also make the first identification of CAstV and group A ARV in broiler chickens in Korea.  相似文献   

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为初步调查SPF鸡感染鹦鹉热嗜性衣原体状况及相关SPF鸡胚源疫苗是否出现污染,本试验通过采集不同日龄的SPF鸡血清70份、SPF种蛋卵黄膜30份,收集市场上销售的SPF鸡胚源疫苗共41支,利用国产间接血凝试剂盒检测抗体,进口免疫荧光试剂盒分别测定其抗体、抗原阳性率,以评价SPF鸡鹦鹉热嗜性衣原体的流行状况和相关疫苗的污染状况。本试验结果显示,SPF鸡血清阳性率分别为31.4%(荧光法)、5.7%(间接血凝法);SPF种蛋阳性率33.3%,SPF鸡胚源疫苗平均阳性率31.7%。SPF鸡已经感染了鹦鹉热嗜性衣原体,且发现经鸡胚卵黄膜而传播病原的新途径,进而造成SPF鸡胚源疫苗出现衣原体污染。因此,加强SPF鸡鹦鹉热嗜性衣原体监测已势在必行。  相似文献   

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Rotavirus particles were identified in the intestinal content of a 35-day-old stunted chicken. The virus was isolated, RNA pattern was analysed and the viral genome segment 6 was sequenced. In particular, the sequence data showed a very close similarity to the chicken rotavirus isolate Ch-1 (99.2% amino acid homology), this is distantly related to all known avian rotaviruses and supports the existence of different VP6 types amongst avian group A rotaviruses.  相似文献   

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禽轮状病毒的分离鉴定及部分特性研究   总被引:5,自引:2,他引:3  
从北京某自然腹泻的鸡场 中成功地分离到一伯禽轮状病毒,并经鸡胚单层肝细胞和MA104细胞进行了传代培养,对其生物学特性作了部分研究,从而为我国禽轮状毒的毒株鉴定、诊断及疫苗等系列研究提供了一定依据。  相似文献   

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To determine the characters of receptors on target cells for avian rotaviruses, the receptors on MA104 cells for the pigeon rotavirus PO-13, the turkey rotaviruses Ty-1 and Ty-3, and the chicken rotavirus Ch-1 were analyzed. Pretreatment of MA104 cells with neuraminidase greatly reduced the infection by all of the four avian rotavirus strains. Binding of the cell-attachment protein, purified VP8 expressed in bacteria, of strain PO-13 to MA104 cells was also inhibited by pretreatment of cells with neuraminidase. These findings suggest that avian rotaviruses primarily utilize sialic acid-containing molecules as receptors on MA 104 cells.  相似文献   

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禽白血病病毒斑点杂交检测方法的建立   总被引:3,自引:0,他引:3  
为建立禽白血病病毒(ALV)斑点杂交检测方法,本研究采用RT-PCR技术扩增ALV群特异性p27抗原基因的部分片段,并以纯化的p27PCR产物为模板,合成地高辛标记探针,以此建立了ALV的斑点杂交检测方法。用该方法对4份疑似感染ALV的现地病鸡组织样品和18枚鸡胚进行检测,结果表明所有样品均为阳性,而且与PCR检测结果的符合率达到100%。该方法具有良好的特异性和敏感性,适应于ALV临床大规模检测。  相似文献   

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为了了解安徽省合肥地区家禽禽流感免疫及带毒情况,2011年4—8月,在合肥地区家禽交易市场,采集不同来源、不同品种家禽的血清和同份棉拭子样品(咽喉、泄殖腔双份)603份(实测589份),分别进行高致病性禽流感免疫抗体和病原学检测,结果表明,全部样品平均免疫抗体合格率为70.80%。对监测结果进行分析发现,不同品种、不同养殖规模、不同月份的高致病性禽流感免疫抗体存在差异,其中,蛋鸡、种鸡高致病性禽流感免疫抗体合格率达80%以上,而肉鸡及水禽的免疫抗体水平则较差;饲养规模在1000羽以上的养禽场家禽高致病性禽流感平均免疫抗体合格率均达到70%以上,而饲养规模在1000羽以下的养禽场和农村散养家禽免疫抗体合格率分别为27.45%和32.84%,尚未达到农业部规定标准;在6月采集的样品,其平均免疫合格率偏低,其他月份差异不显著。病原学检测结果表明,全部检测样品高致病性禽流感病原检测结果均为阴性。  相似文献   

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Zhang A  Jin M  Liu Ff  Guo X  Hu Q  Han L  Tan Y  Chen H 《Avian diseases》2006,50(3):325-330
Rapid detection of avian influenza virus (AIV) infection is critical for control of avian influenza (AI) and for reducing the risk of pandemic human influenza. A double antibody sandwich enzyme-linked immunosorbent assay (DAS-ELISA) was developed for this purpose. The method employed a monoclonal antibody (MAb) as the capture antibody and rabbit polyclonal IgG labeled with horseradish peroxidase as the detector antibody, and both antibodies were against type-specific influenza A nucleoprotein (NP). The DAS-ELISA could detect minimally 2.5 ng of influenza viral protein in virus preparations treated with Triton X-100, which is equvilent to 2.5 x 10(2) EID50 virus particles. This DAS-ELISA could detect all 15n AIV subtypes (H1-H15) and did not cross react with other avian pathogens tested. The DAS-ELISA were directly compared with virus isolation (VI) in embryonated chicken eggs, the current standard of influenza virus detection, for 805 chicken samples. The DAS-ELISA results correlated with VI results for 98.6% of these samples, indicating a sensitivity of 97.4% and specificity of 100%. The method was further tested with H5N1 and H9N2 AIV experimentally infected chickens, ducks, and pigeons, as well as field samples obtained from central China in 2005. The DAS-ELISA method has demonstrated application potential as an AIV screening tool and as a supplement for virus isolation in Asia.  相似文献   

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Cohort studies were conducted on 29 pigs from 3 villages in the Highlands of Papua New Guinea. Animals ranged in age from 9 d to 5 m old. Three hundred and twenty nine faecal samples were collected from individual pigs followed over 3 to 6 w periods, and were examined for group A rotavirus antigen by ELISA, and rotaviral genomic RNA by polyacrylamide gel electrophoresis (PAGE). Electron microscopy was also conducted on selected samples. Group A rotavirus was detected in the faeces of 16 pigs with infected individuals coming from all villages. Non-group A rotavirus resembling group C was found in faeces from pigs from 2 villages. All of the group A rotaviruses examined had the same electrophoretype and this was distinct from that of the common type infecting humans in the area at the time of the study. None of the group A positive samples reacted with monoclonal antisera specific for human group A rotaviruses of serotypes 1, 2, 3, 4, or 8. The non-group A rotaviruses also all had identical electrophoretypes. In contrast to previous findings in intensive piggeries, rotavirus infection did not occur in all young pigs and was not limited to young animals under 2 m of age. Infected pigs varied in age from 12 days to 20 weeks of age. This pattern of infection was attributed to the non-intensive husbandry situations in the villages, with less opportunity for transmission to occur than in intensive piggeries.  相似文献   

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Agar gel immunodiffusion (AGID) and counter-immunoelectrophoresis (CIEP), complement fixation (CF), radio-immunoassay (RIA), haemagglutination (HA) and haemagglutination inhibition (HI) tests were compared in their efficiency for the detection of bovine rotavirus antigens and antibodies. As a test for antigen using hyperimmune serum, CIEP was found to have advantages over AGID by being more rapid as well as approximately four times more sensitive regardless of whether the antigen was of faecal or tissue culture origin. The CF test was more sensitive than either of the immunodiffusion procedures studied for antigen detection, but was more tedious to perform and of limited use as some faecal samples exhibited anti-complementary activity. For measurement of rotavirus antibody the radio-immunoassay (RIA) was the most sensitive technique and the CIEP least sensitive. Using the RIA a limited survey of cattle demonstrated that approximately 75% of the animals tested possessed specific antibody to rotavirus.  相似文献   

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Detection and survival of group A rotavirus in a piggery   总被引:1,自引:0,他引:1  
Samples of dust, faeces and effluent were collected from a piggery and examined for group A rotavirus, using a commercial ELISA test, electron microscopy and inoculation of MA-104 cells. Rotavirus antigen was demonstrated in samples collected from farrowing and weaner rooms but not from fattener and sow houses. Rotavirus antigen was also detected in samples collected from a weaner room which had been free of piglets for three months. A cytopathic porcine rotavirus (British isolate SW20/21) was kept at room temperature for four months; it survived with titres reduced by 2 log10. These observations suggest that the environment of commercial piggeries is an important source of rotaviral infection for young piglets.  相似文献   

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应用识别不同表位的鸡白细胞介素18成熟蛋白(Mature chicken interleukin-18,mChIL-18)的2株单克隆抗体(mAb)1G9和2E6,建立检测mChlL-18的双抗体夹心ELISA,并利用此方法对禽网状内皮组织增生症病毒(Reticuloendotheliosis virus,REV)人工感染SPF鸡体内mChIL-18的分泌水平进行检测。结果显示,捕获抗体的最佳质量浓度为8mg/L,检测抗体的工作效价为1:800,待检样品的最佳稀释度为1:400,检测敏感度可达31.5ng/L,与其他细胞因子等抗原蛋白无交叉反应;跟对照组相比,REV感染鸡体内mChIL-18的表达量在7、14、21、28、35、42、49d均呈现升高,但只有14日龄时表现差异显著(P〈0.05)。结果表明,本试验成功建立了ChIL-18的双抗体夹心ELISA,为鸡传染病的细胞免疫学研究提供了可靠方法。  相似文献   

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Rotaviruses are the main agents responsible for diarrhea in different animal species and for infantile gastroenteritis. These viruses have been isolated from various avian species and have often been associated with poult enteritis and mortality syndrome. Nevertheless, the knowledge of rotavirus infection in turkeys is scarce. Six group A rotavirus strains obtained from pooled enteric contents of diarrheic turkeys were isolated in MA-104 cell culture and typed as G(6)P(1), a typical bovine rotavirus genotype. Additionally, the electropherotypes showed a migration pattern identical to the Nebraska calf diarrhea virus, and the complete NSP4 gene phylogeny showed that all six strains segregated in the genotype E2. Taken together, these results point toward a cattle-to-turkey rotavirus transmission. As a conclusion, bovine-origin rotavirus can be found in turkeys, and this transmission route must now be considered for the improvement of the health status in turkey farms.  相似文献   

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Three precipitin reactions associated with bovine rotavirus infection were demonstrable by immunodiffusion. One of the reactions has been utilized in a diagnostic test for the detection of rotavirus in faeces, or specific antibody to rotavirus group antigen in serum or faeces. The test, based on bovine materials, appeared to be group-specific and effective in demonstrating rotaviral antigen or antibody in other species of animals, including human beings. The procedure was as efficient as electron microscopy in detecting evidence of rotavirus in faeces of calves and a range of other species.  相似文献   

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