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The ability of the portable Cepheid SmartCycler real-time PCR machine to detect foot-and-mouth disease (FMD) virus sensitively and accurately was evaluated by comparing the results of the analyses of nasal swab and serum samples from experimentally infected animals with those obtained from the real-time PCR assay currently in use in the laboratory. The results indicated that the ability of the machine to detect viral RNA is greatly affected by the PCR reagents used for the assay. When it was used with PCR beads it was unable to detect weakly positive samples, but when TaqMan core reagents were used for the assay, its sensitivity was significantly increased. The machine could be used for the laboratory-based detection of FMD; however, as with all assays, significant optimisation of assay conditions as well as solid validation of the technique is required.  相似文献   

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This study was carried out to determine serum protein profiles in naturally infected sheep with foot-and-mouth disease virus (FMDV). The study material consisted of twelve healthy and 36 sheep with foot-and-mouth disease (FMD). FMD had been diagnosed on the basis of clinical findings and results of serological examination. Serotypes serologically detected in the FMDV-infected sheep were as follows: O (n = 11), A (n = 8) and mixed infection with serotypes O, A and Asia-1 (n = 17).The total protein, albumin and globulin concentrations as well as Albumin/Globulin ratio were slightly different among the groups (P < 0.05). Three protein bands of 66 kDa, 45 kDa and 20 kDa were remarkable. Moderate differences were determined between healthy and infected sheep for proportion of distribution in serum proteins. In conclusion, serum protein concentrations and serum protein profiles were slightly changed and no specific serum protein profile occurred in sheep infected with either O or A or in sheep mixed infected with the O and A and Asia-1 serotypes of FMDV compared to healthy ones.  相似文献   

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Quantitative analysis of the duration of foot-and-mouth disease virus (FMDV) RNA in tissues was carried out in pigs experimentally infected with FMDV O UKG 34/2001 and O SKR 1/2000. The results showed that the viral RNA was still detectable in cervical lymph nodes, mandibular lymph nodes and tonsils collected from both inoculated and contact pigs at 28 days post infection. There was no detectable viral RNA in the soft palate or pharynx, which are thought to be tissue sites for viral persistence in cattle. Further study is needed to clarify whether this difference has significance in terms of viral clearance in pigs.  相似文献   

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口蹄疫病毒能引起牛、羊等偶蹄动物发生高度接触性的传染病口蹄疲,该病常常影响着全球畜牧业的发展.FMDV是小RNA病毒科口蹄疫病毒属的成员,口蹄疫病毒为单股正链RNA病毒,病毒基因组全长约8.5 kb,基因组分为5'非编码区、3'非编码区和一个开放阅读框(ORF).基因组的中部是一大的开放阅读框,编码一多聚蛋白,多聚蛋白在翻译的同时,经二级裂解后,形成3种病毒结构蛋白(VP0,VP3和VP1)和8种非结构蛋白(L,2A,2B,2C,3A,3B,3C和3D).其中3C全长639 bp,编码213个氨基酸.3C蛋白酶是小RNA病毒的共同裂解酶,在多聚蛋白成熟过程中起着极为重要的作用,且在抗病毒药物打靶方面具有一定研究价值.因此,对3C蛋白酶的结构及功能研究进展进行综述很有必要.  相似文献   

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根据GenBank中O型和Asia1型口蹄疫病毒(Foot-and-mouth disease virus,FMDV)的vp3、vp1和2A基因序列,并与其它血清型FMDV的对应基因序列进行比较,设计用于扩增O型和Asia1型FMDV vp1基因的特异性引物,建立O型和Asia1型FMDV RT-PCR鉴别诊断方法。本方法首先用通用型引物进行RT-PCR,确定是否为FMDV感染,然后用特异性引物鉴别O型或Asia1型FMDV的感染。用vp1基因序列分析进行符合性试验,验证了该方法所具有的特异性和敏感性。本方法可用于O型和Asia1型FMD的快速诊断及流行病学调查。  相似文献   

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鉴于现有疫苗和抗病毒药物在治疗口蹄疫方面存在的局限性,寻找新型抗病毒策略势在必行,而RNA干扰正是一种有益的尝试。RNA干扰通过沉默哺乳动物细胞中的基因表达,可有效的阻断口蹄疫病毒在宿主体内的复制。论文以口蹄疫病毒引起宿主细胞病变、RNA干扰的机制及RNA干扰抑制FMDV在宿主细胞内复制为内容,探讨如何更加有效地防控口蹄疫。  相似文献   

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A panel of 36 sera has been assembled from experimental cattle that had been infected by inoculation or contact exposure with 4 serotypes of foot-and-mouth disease virus (FMDV) with or without prior vaccination. Virus replication and persistence had been characterized in all of the animals. The proportion of the sera scored positive by 5 tests for antibodies to the nonstructural proteins of FMDV varied, suggesting that the panel can discriminate between the sensitivity with which such tests are able to identify infected cattle. Use of this panel will help in assessment of new tests and quality control of existing methods.  相似文献   

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Foot-and-mouth disease (FMD) is an acute,febrile and highly contagious animal disease caused by foot-and-mouth disease virus (FMDV),and has been recognized as the most important constraint to international trade in animals and animal products.An outstanding feature for FMDV infection is that the FMDV infected animals may remain as a carrier state,some of the animals exposed to FMDV may have a long term asymptomatic infection.This article will review the advance of FMDV in the following aspects,epidemiology,etiology and pathogenesis.  相似文献   

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口蹄疫是由口蹄疫病毒引起的主要侵袭偶蹄动物的一种急性热性高度接触性传染病。口蹄疫病毒为微RNA病毒科口蹄疫病毒属成员,存在7个不同血清型,病毒VP1蛋白抗原性差异是病毒血清型划分依据,而其编码基因(1D)核苷酸序列差异是同型病毒拓扑型(Topotype)或基因型鉴别依据。采用O/A/C/Asia-1多重RT-PCR技术,对2006年自云南边境地区采集的120份动物组织样品,进行口蹄疫病原监测,检出O型口蹄疫病毒阳性样品15份。对阳性样品中病毒VP1基因全序列进行扩增、纯化后,克隆至pMD18-T载体测序,并与已知代表性毒株进行比对及系统发育分析。结果发现:云南边境O型口蹄疫病毒阳性样品VP1基因核苷酸序列同源性介于77.3%~98.7%,可划分为3个不同的拓扑型或基因型:中东-南亚型(ME-SA)或泛亚型(PAN-Asia)、古典中国型(Cathay)、东南亚型(SEA)。部分样品VP1蛋白表位43位、154位关键性氨基酸位点存在变异。  相似文献   

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口蹄疫(foot-and-mouth disease,FMD)是由口蹄疫病毒(foot-and-mouth disease virus,FMDV)引起的一种急性、热性、高度接触传染性动物疫病,是全球范围内家畜及其产品贸易最大的羁绊。FMDV通过逃避宿主的免疫监视建立持续性感染,使患畜持续向外界排毒,成为传染源。作者查阅了近几年FMDV的国内外研究进展,对其流行病学、病原学及致病机理进行了概述。  相似文献   

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A quarantine period for potentially contaminated personnel can be used to reduce the risk of transfer of foot-and-mouth disease virus (FMDV) from infected to susceptible premises. This is set at 72 hours in the UK, on the basis of results from laboratory studies and field observations. Previous analysis of FMDV carriage within human nasal cavities has relied upon virus isolation by culture in susceptible cells. This study, involving 51 people, evaluated a PCR method, which detected viral genomic material within 35 nasal swabs taken from personnel after up to eight hours exposure to infected animals. Only one of 23 people who was PCR-positive immediately after exposure to FMDV-infected animals remained positive the following day, indicating a low risk of prolonged carriage of virus in the nasal cavities.  相似文献   

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The presence of serum antibodies for nonstructural proteins of the foot-and-mouth disease virus (FMDV) can differentiate FMDV-infected animals from vaccinated animals. In this study, a sandwich ELISA was developed for rapid detection of the foot-and-mouth disease (FMD) antibodies; it was based on an Escherichia coli-expressed, highly conserved region of the 3ABC nonstructural protein of the FMDV O/TW/99 strain and a monoclonal antibody derived from the expressed protein. The diagnostic sensitivity of the assay was 98.4%, and the diagnostic specificity was 100% for na?ve and vaccinated pigs; the detection ability of the assay was comparable those of the PrioCHECK and UBI kits. There was 97.5, 93.4 and 66.6% agreement between the results obtained from our ELISA and those obtained from the PrioCHECK, UBI and CHEKIT kits, respectively. The kappa statistics were 0.95, 0.87 and 0.37, respectively. Moreover, antibodies for nonstructural proteins of the serotypes A, C, Asia 1, SAT 1, SAT 2 and SAT 3 were also detected in bovine sera. Furthermore, the absence of cross-reactions generated by different antibody titers against the swine vesicular disease virus and vesicular stomatitis virus (VSV) was also highlighted in this assay's specificity.  相似文献   

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A novel proximity ligation assay (PLA) using a pan-serotype reactive monoclonal antibody was developed and evaluated for the detection of foot-and-mouth disease virus (FMDV) in clinical samples collected from field cases of disease. The FMDV-specific PLA was found to be 100 times more sensitive for virus detection than the commonly used antigen capture-ELISA (AgELISA). As few as five TCID50 were detected in individual assays, which was comparable with the analytical sensitivity of real-time RT-PCR. Although this assay was capable of detecting diverse isolates from all seven FMDV serotypes, the diagnostic sensitivity of the PLA assay was lower than real-time RT-PCR mainly due to a failure to detect some SAT 1, SAT 2 and SAT 3 FMDV strains. In conclusion, this new PLA format has high analytical sensitivity for the detection of FMDV in clinical samples and may prove valuable as a rapid and simple tool for use in FMD diagnosis.  相似文献   

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In order to use nucleotide sequencing for foot-and-mouth disease virus (FMDV) diagnostic subtyping, it is necessary to shorten the time required for preparation of suitable templates. The time required for analysis was reduced by use of the viral RNA present in the total RNA extract of tissue from infected cattle as a template in the Sanger sequencing reaction. Results are now available within 3 days. The sequences determined encode capsid protein VP1 and therefore major neutralization epitopes. Such a sequence of FMDV O1Kaufbeuren, cultured in the animal, was compared with those of tissue-cultured viruses. They did not differ. It was concluded that a change of virus culture conditions does not necessarily account for antigenic variation.  相似文献   

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