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兔病毒性出血症是由兔出血症病毒（Rabbit hemorrhagic disease virus，RHDV）引起的一种急性、高度致死性传染病，病死率高达100％。近年来，由于该病毒血凝性、抗原性的变异，病毒感染宿主增多，不仅对该病的防控造成了一定的威胁，更造成了严重的经济损失。国内外研究学者对该病毒的基因组结构、基因组编码的产物和功能，病毒的宿主和受体以及病毒变异等相关进行了大量深入的研究和分析，为该病毒的防控和治疗提供了技术基础。本文就RHDV遗传与变异的分子基础作一综述，希望能为深入了解该病毒，以及日后的防控提供一定的理论支持。 相似文献
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Luzia Teixeira Raquel M. Marques Artur P. Águas Paula G. Ferreira 《Research in veterinary science》2011,91(1):164-166
Rabbit Haemorrhagic Disease Virus (RHDV), a member of the Caliciviridae family, is the etiologic agent of Rabbit Haemorrhagic Disease (RHD); this viral disease is highly contagious and kills more than 90% of infected adult rabbits. Research on experimental calicivirus infection uses inocula obtained from livers of rabbits dying from calicivirus infection. This implies that caliciviruses have to be purified from liver homogenates. Current methods to isolate caliciviruses from rabbit livers are time consuming. We propose here a new procedure for fast purification of rabbit caliciviruses from liver homogenates that uses centrifugation through an iodixanol gradient. This method offers in approximately 2 h a sample with a high degree of calicivirus purity, as shown by its biochemical and immunocytochemistry analysis, which is also able to kill adult rabbits from RHD within 48 h of inoculation. 相似文献
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AIM: To test for antibodies to rabbit haemorrhagic disease (RHD) virus (RHDV) in sera from mammals and birds associated with rabbit populations infected with RHDV. METHODS: Sera from feral and domestic cats, feral ferrets, stoats, hedgehogs, hares, harrier hawks, and black-backed gulls were taken (apart from some of the hares) from areas in New Zealand where RHD was active among rabbit populations. The presence of antibodies to RHD was investigated using a competition enzyme-linked immunosorbent assay (ELISA). RESULTS: Some individual animals of all species were seropositive. Thirty eight of 71 feral cats, but only 1/80 domestic cats were seropositive at a 1:40 dilution. The latter had not been exposed to RHDV. Also reactive in the ELISA were 2/8 stoats; 11/115 ferrets, with significantly more females having antibodies than males; 4/73 hedgehogs; 2/18 hawks, and 1/30 gulls. Three of 66 hares, comprising 3/14 from one population, were seropositive. CONCLUSIONS: Apart from the hares, all these species are known to prey upon rabbits or scavenge their carcasses, a possible means of exposure to RHDV. The possibility that the positive test reactions were due to cross-reactions with other caliciviruses cannot be ruled out, especially for the hares. Nor could the study differentiate whether the positive results were due to an antigenic reaction to ingestion of RHDV, as suggested by overseas work, or to infection of new species by RHDV. These possibilities are being investigated further. 相似文献
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新型兔出血症病毒(RHDV2)在全球范围流行并已在我国出现,给养兔业带来巨大威胁。为了解RHDV2与经典兔出血症病毒(RHDV)的异同,从病原学、流行病学、临床症状、致病机理、诊断与防控等方面,阐述了两种病原的不同及相关研究的最新进展。对比发现:RHDV2与经典RHDV亲缘关系较远,抗原表位也存在明显差异,属于不同的病毒分支;经典RHDV主要感染成年家兔,而RHDV2对不同品种和年龄的兔均易感;经典RHDV感染常表现为急性症状,而新型RHDV感染以亚急性和慢性病症为主。目前针对RHDV2的检测和疫苗研究还相对滞后。鉴于RHDV2与经典RHDV之间免疫交叉性较弱,免疫预防上存在较大空白,下一步应加大RHDV2毒株结构功能研究,加快疫苗开发,做好可疑病例处置和流行病学研究,及时掌握我国疫情动态。本文为我国兔病毒性出血症的预防与控制提供了参考。 相似文献
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Serological cross reactivity between the virulent rabbit haemorrhagic disease virus (RHDV) and the closely related but non-pathogenic rabbit calicivirus (RCV) makes it difficult to study the epidemiology of each virus and the interaction between them when both viruses co-circulate in wild rabbit populations. ELISA methods for the diagnosis of RHDV infection are well characterized, but no specific serological tests for RCV have been developed. Following the characterization of Australian non-pathogenic RCV-A1 strains, we used virus-like-particles (VLPs) and anti-RCV-A1 specific antibodies to establish a set of isotype ELISAs for detection of IgG, IgA and IgM in rabbit sera and secretory mucosal IgA in rectal swabs, and two competition ELISAs. These assays were used to discriminate between anti-RCV-A1 and anti-RHDV antibodies in rabbits. The isotype ELISAs were highly sensitive for detection of anti-RCV-A1 antibodies, but varying levels of cross reactivity from anti-RHDV antibodies occurred in the isotype ELISAs and one competition ELISA. However, the second competition ELISA specifically detected antibodies to RCV-A1 and showed no cross reactivity to anti-RHDV sera. These ELISAs provide important tools to monitor RCV-A1 infection when it occurs alone, and to discriminate between RHDV and RCV-A1 infection when they occur in the same rabbit population. When used in parallel with RHDV serology, they could be used to monitor the dynamics of these two closely related but pathogenically distinct viruses in wild and domestic rabbit populations. 相似文献
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根据GenBank已发表的多株RHDV全基因组序列，设计3对扩增RHDVCD株的特异性引物，扩增了RHDV CD株包含全序列的3个片段，并分别克隆到pMD18-T载体上。根据引物上唯一的交叉酶切位点，把三个克隆片段同时克隆到pUC18载体上，获得了RHDV CD株全基因组克隆，测定了全基因组序列（GenBank accession number AY523410）。进行了CD株全序列与GenBank上已发布的其他6株全序列比较及基因进化树分析。发现除CD株外，其余6株（西班牙AST-89株、法国SD株、意大利BS89株、德国分离1株、德国分离2株、澳大利亚Czech V351株）之间的同源性均很高，在93．9％～100％之间，而CD株与其他6株之间的同源性均较低，在89．0％～89．6％之间。德国的2个分离株序列完全相同，应为同一毒株：进化树分析表明，此7株病毒可形成2个明显的分支，中国CD株形成一个分支，其余欧洲、澳大利亚分离株开成一个分支，具有明显地域差异特征． 相似文献
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为建立一种快速的兔病毒性出血症病毒(rabbit hemorrhagic disease virus,RHDV)病原检测方法,本研究根据GenBank上登录的RHDV VP60基因序列,设计合成内外2对引物,优化PCR反应条件,建立了检测RHDV的巢式RT-PCR方法。该方法对兔轮状病毒、仙台病毒、健康兔肝脏组织的扩增结果均为阴性;该方法第1次扩增的敏感性是10 ng,第2次扩增的敏感性是0.1 ng,第2次比第1次扩增的敏感性高100倍。建立的巢式RT-PCR方法具有特异性强、敏感性高、重复性好等优点,可以准确快速检测出极低含量的RHDV,将为兔病毒性出血症的病原检测及分子流行病学调查等提供一种快速、简单、高效、特异、灵敏的检测方法。 相似文献
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Juan Bárcena Beatriz Guerra Iván Angulo Julia González Félix Valcárcel Carlos P. Mata José R. Castón Esther Blanco Alí Alejo 《Veterinary research》2015,46(1)
In 2010 a new Lagovirus related to rabbit haemorrhagic disease virus (RHDV) emerged in France and has since rapidly spread throughout domestic and wild rabbit populations of several European countries. The new virus, termed RHDV2, exhibits distinctive genetic, antigenic and pathogenic features. Notably, RHDV2 kills rabbits previously vaccinated with RHDV vaccines. Here we report for the first time the generation and characterization of RHDV2-specific virus-like particles (VLPs). Our results further confirmed the differential antigenic properties exhibited by RHDV and RHDV2, highlighting the need of using RHDV2-specific diagnostic assays to monitor the spread of this new virus. 相似文献
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Fernández E Toledo JR Chiong M Parra F Rodríguez E Montero C Méndez L Capucci L Farnós O 《Veterinary immunology and immunopathology》2011,142(3-4):179-188
Rabbit hemorrhagic disease virus (RHDV) is the etiological agent of a lethal and contagious disease of rabbits that remains as a serious problem worldwide. As this virus does not replicate in cell culture systems, the capsid protein gene has been expressed in heterologous hosts or inserted in replication-competent viruses in order to obtain non-conventional RHDV vaccines. However, due to technological or safety issues, current RHDV vaccines are still prepared from organs of infected rabbits. In this work, two human type 5 derived replication-defective adenoviruses encoding the rabbit hemorrhagic disease virus VP60 capsid protein were constructed. The recombinant protein was expressed as a multimer in mouse and rabbit cell lines at levels that ranged from approximately 120 to 160 mg/L of culture. Mice intravenously or subcutaneously inoculated with a single 10(8) gene transfer units (GTU) dose of the AdVP60 vector (designed for VP60 intracellular expression) seroconverted at days 7 and 14 post-immunization, respectively. This vector generated a stronger response than that obtained with a second vector (AdVP60sec) designed for VP60 secretion. Rabbits were then immunized by parenteral or mucosal routes with a single 10(9)GTU dose of the AdVP60 and the antibody response was evaluated using a competition ELISA specific for RHDV or RHDVa. Protective hemagglutination inhibition (HI) titers were also promptly detected and IgG antibodies corresponding with inhibition percentages over 85% persisted up to one year in all rabbits, independently of the immunization route employed. These levels were similar to those elicited with inactivated RHDV or with VP60 obtained from yeast or insect cells. IgA specific antibodies were only found in saliva of rabbits immunized by intranasal instillation. The feasibility of VP60 production and vaccination of rabbits with replication-defective adenoviral vectors was demonstrated. 相似文献
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Di Martino B Martella V Di Profio F Ceci C Marsilio F 《Veterinary microbiology》2011,149(1-2):221-224
St-Valerien-like viruses are newly recognized porcine caliciviruses recently detected in Canadian swine farms. These viruses that appeared genetically closest to the Tulane non-human primate virus and noroviruses (NoVs), may represent members of a potential new genera within the Caliciviridae family. To date, there are no information on the epidemiology of these novel caliciviruses in other continents. In this study, 264 faecal samples collected from asymptomatic adult pigs were screened by RT-PCR using St-Valerien-like specific primers. Porcine caliciviruses, resembling St-Valerien-like viruses were identified in five animals. The 3' end of the genome of one of these strains, 25A/ITA, was determined, revealing close genetic relatedness to the newly discovered St-Valerien-like caliciviruses, identified in swine in Canada. The findings of this investigation demonstrate that St-Valerien-like viruses are not geographically restricted, since they were identified outside the North American continent where there were first signalled. 相似文献
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试验旨在分离兔出血症病毒（rabbit hemorrhagic disease virus,RHDV）镇江株,分析其遗传进化变异,并表达具有良好活性的重组VP60蛋白。通过排除细菌感染、血凝（HA）和血凝抑制（HI）检测、动物攻毒试验与病毒传代、LD50测定等方法,自江苏省镇江市某兔场发病死亡动物肝脏组织样品中分离病原并鉴定;RT-PCR方法获得VP60基因,通过分析VP60基因核苷酸及氨基酸序列研究其遗传进化;将VP60基因与pCold-Sumo载体连接,构建低温诱导、融合Sumo标签原核表达载体,15℃、IPTG诱导表达重组VP60蛋白并对表达产物进行反应原性鉴定。结果显示,分离鉴定获得RHDV ZJ2015毒株,该毒株能凝集人"O"型红血球,HA效价为11log2,其血凝性能被RHDV （AV33）抗血清抑制,该毒株的LD50为10-6.38/mL,具有较强的毒力;RT-PCR扩增得到大小约为1 740 bp的特异性条带,系统进化树分析显示,该毒株属于RHDV1抗原遗传变异株（RHDVa）,与RHDV1和RHDV2 VP60基因核苷酸序列同源性分别为89.4%~97.6%和81.1%~81.5%,氨基酸序列同源性分别为93.8%~98.3%和87.4%~87.6%。构建的低温原核融合表达质粒pCold-VP60在大肠杆菌Rosetta （DE3）中成功表达,通过SDS-PAGE及Western blotting分析,在分子质量74 ku处有特异性表达蛋白条带,且与抗血清发生特异性抗原抗体结合反应,说明重组蛋白具有良好的反应原性。本研究为开展兔病毒性出血症流行病学研究、开发新型重组疫苗与诊断试剂提供参考依据。 相似文献
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Muller A Freitas J Silva E Le Gall-Reculé G Zwingelstein F Abrantes J Esteves PJ Alves PC van der Loo W Kolodziejek J Nowotny N Thompson G 《Veterinary microbiology》2009,135(3-4):368-373
To date information on rabbit haemorrhagic disease virus (RHDV) in Spain and Portugal has been scarce, although the disease is endemic and continues to have a considerable impact on species conservation and hunting industry. We analysed RHDVs obtained between 1994 and 2007 at different geographic locations in Portugal (40 samples), Spain (3 samples) and France (4 samples) from wild European rabbits (Oryctolagus cuniculus) that succumbed to the disease. Phylogenetic analyses based on partial VP60 gene sequences allowed a grouping of these RHDVs into three groups, termed "Iberian" Groups IB1, IB2 and IB3. Interestingly, these three Iberian groups clustered separately, though not far from earlier RHDVs of Genogroup 1 (containing e.g., strain "AST89"), but clearly distinct from globally described RHDV strains of Genogroups 2-6. This result, supported by a bootstrap value of 76%, gives rise to the hypothesis that the virus evolved independently since its introduction to wild rabbit populations on the Iberian Peninsula, with the Pyrenees acting as a natural barrier to rabbit and hence to virus dispersal. No differences were observed in RHDV sequences obtained from geographic regions where the rabbit subspecies O. c. algirus prevails compared with those obtained from O. c. cuniculus. 相似文献
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Four years after the release of Rabbit haemorrhagic disease virus (RHDV) in New Zealand, we conducted a mail survey of farmers to ascertain their attitudes and practices about rabbit control. A multistage sampling frame (stratified by rabbit-proneness and farm type) was used to select 828 farms in eight geographical regions. The useable response proportion of the survey was 69.3%, and 21% of respondents considered rabbits to be a problem on their farms. Although practices for rabbit control had changed from 1995 to 2001, shooting (practised by 85% of respondents) remained the predominant method used (albeit less frequently than in 1995). Ten percent of farmers used RHDV baiting; of those, 90% released the virus relatively infrequently. Farmers perceived shooting to be the most-humane and environmentally safe method, while RHDV was perceived to be the most effective. Perception of the level of competition for grazing between rabbits and livestock was the factor most-strongly associated with the use of shooting and RHDV. Most (60%) respondents considered the introduction of RHDV to have been beneficial. 相似文献
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为鉴定兔出血症病毒(RHDV)在感染兔肝脏过程中与肝细胞膜表面相互作用的靶蛋白,本研究利用SMART文库构建技术构建了兔肝脏细胞的真核表达cDNA重组质粒文库,将其转染SP2/0细胞,经嘌呤霉素筛选表达兔肝细胞蛋白的阳性SP2/0细胞.以RHDV-VP60蛋白作为固相包被抗原,经筛选,获得能够与VP60蛋白相互结合的表达性阳性SP2/0细胞克隆.测序表明26个克隆与兔的一些功能性蛋白有较高的同源性,其中包括代谢酶类13个、免疫信号通路受体蛋白5个以及其他细胞膜蛋白等.本研究鉴定的与RHDV VP60具有结合性的细胞蛋白为RHDV感染机制的研究奠定了基础. 相似文献
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AIM: To estimate over a 3-year period following the first release of rabbit haemorrhagic disease virus (RHDV) the prevalence of rabbit haemorrhagic disease (RHD) and the abundance of rabbits (Oryctolagus cuniculus) in an area that historically had low rabbit densities. METHODS: Three farms grazing predominantly sheep and beef cattle, located close together and with low initial rabbit densities, were selected for study. RHDV had been deliberately released on all farms in December 1997. Farms were visited 2-3 times per year between June 1998 and April 2001. At each visit, rabbits were shot with the aid of spotlights at night and blood samples were collected for detection of RHDV antibodies. Rabbit carcasses were necropsied and the age of the animals was determined. Rabbit abundance on each property was measured throughout the study using spotlight night counts. Logistic regression was used to identify factors associated with the risk of carcasses being seropositive for RHDV. RESULTS: Rabbit density differed initially between farms (8.2, 9.9, 2.3 rabbits per spotlight km in June 1998), and declined on all three properties over time (1.2, 2.4, 1.1 rabbits per spotlight km in November 2000). Highest antibody titres to RHDV were initially evident on the farm on which rabbits were most abundant. The average prevalence of seropositive rabbits overall was 21% (95% CI=15-28%). Female rabbits tended to be less likely to be seropositive for RHDV than males (OR=0.47; 95% CI=0.21-1.02). The odds of becoming seropositive were reduced for rabbits born in the breeding season of 1999-2000 (OR=0.17; 95% CI=0.05-0.64). CONCLUSIONS: The temporal pattern of outbreaks measured by peaks of seroprevalence differed between closely-spaced farms when they had different rabbit densities, but were similar when rabbit densities were similar. Microclimate and vegetation influencing abundance of insect vectors for RHDV and intrinsic population-related factors like rabbit breeding behaviour are also likely to be involved in local patterns of spread. 相似文献
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AIM: To estimate over a 3-year period following the first release of rabbit haemorrhagic disease virus (RHDV) the prevalence of rabbit haemorrhagic disease (RHD) and the abundance of rabbits (Oryctolagus cuniculus) in an area that historically had low rabbit densities. METHODS: Three farms grazing predominantly sheep and beef cattle, located close together and with low initial rabbit densities, were selected for study. RHDV had been deliberately released on all farms in December 1997. Farms were visited 2–3 times per year between June 1998 and April 2001. At each visit, rabbits were shot with the aid of spotlights at night and blood samples were collected for detection of RHDV antibodies. Rabbit carcasses were necropsied and the age of the animals was determined. Rabbit abundance on each property was measured throughout the study using spotlight night counts. Logistic regression was used to identify factors associated with the risk of carcasses being seropositive for RHDV. RESULTS: Rabbit density differed initially between farms (8.2, 9.9, 2.3 rabbits per spotlight km in June 1998), and declined on all three properties over time (1.2, 2.4, 1.1 rabbits per spotlight km in November 2000). Highest antibody titres to RHDV were initially evident on the farm on which rabbits were most abundant. The average prevalence of seropositive rabbits overall was 21% (95% CI=15–28%). Female rabbits tended to be less likely to be seropositive for RHDV than males (OR=0.47; 95% CI=0.21–1.02). The odds of becoming seropositive were reduced for rabbits born in the breeding season of 1999–2000 (OR=0.17; 95% CI=0.05–0.64). CONCLUSIONS: The temporal pattern of outbreaks measured by peaks of seroprevalence differed between closely-spaced farms when they had different rabbit densities, but were similar when rabbit densities were similar. Microclimate and vegetation influencing abundance of insect vectors for RHDV and intrinsic population-related factors like rabbit breeding behaviour are also likely to be involved in local patterns of spread. 相似文献
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本试验旨在鉴定四川金堂某兔场疑似兔出血症病毒2型（RHDV2）感染疫情的病原,并分析病兔的病理组织学变化。利用血凝试验和RT-PCR检测病死兔内脏组织中的病原,取病变组织制作病理切片,观察分析各组织的病理组织学变化,同时应用病兔肝脏悬液感染幼兔,分析该毒株的致病力。血凝试验结果显示,所采集病死兔肝脏样品能凝集人"O"型血红细胞;RT-PCR扩增及测序结果显示,多对引物均能从样品中扩增出RHDV2特异性条带;病理组织学观察结果显示,病兔多脏器严重出血、肿胀,淋巴细胞和中性粒细胞大量浸润,气管黏膜、肝脏、肺脏出血尤为严重;动物试验结果显示,该毒株毒力较强,含毒肝脏悬液能在24 h内迅速致死幼兔。本研究经临床诊断、核酸检测及测序证实了此次疫情确由RHDV2感染引起,动物试验和病理组织学观察表明该毒株毒力较强,可引发脏器严重出血,造成病兔急性死亡,RHDV2的出现提示病毒的跨境传播情况不容乐观,应引起更大的重视。 相似文献