共查询到17条相似文献，搜索用时 187 毫秒
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《广东畜牧兽医科技》2015,(5)
从山东东营某兔场送检的疑似兔病毒性出血症(Rabbit Hemorrhagic Disease,RHD)病料中分离到1株病毒(DY株)。通过血凝性及RT-PCR鉴定,以及毒力和免疫原性测定。结果显示:分离株DY株为兔出血症病毒(Rabbit Hemorrhagic Disease Virus,RHDV),对人O型红细胞具有高度凝集性,血凝效价为12log2,RHDV抗血清可特异性抑制这种凝集作用;RT-PCR扩增出RHDV VP60的201 bp基因片段;RHDV DY株对家兔的LD50为10-6.5/m L,是一株对家兔具有高致病力的强毒株。取第5代RHDV DY株制备灭活疫苗免疫试验兔,免疫后第14d攻毒,对RHDV的保护率达100%。 相似文献
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为了对南部县某兔场疑似兔出血症病毒(RHDV)NB毒株进行鉴定,试验采用血凝(HA)及血凝抑制(HI)试验、家兔接种试验、家兔免疫攻毒试验、VP60基因的同源性比对。结果显示:NB毒株能凝集人"O"型红细胞,HA效价为12 log2,其血凝性能被RHDV疫苗毒株AV33株的抗血清抑制;NB毒株注射健康非免家兔,家兔在48h内死亡,具有典型的兔病毒性出血症(RHD)的临床症状和病理变化;RHDV(AV33)组织灭活疫苗免疫家兔后,家兔能抵抗NB毒株的攻击;NB毒株与AV33毒株的VP60基因同源性为96.12%,氨基酸序列同源性为97.59%。 相似文献
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为确定发病兔场兔的死亡病因并对其病原进行鉴定分析,经临床诊断、病理剖检、病毒分离、纯净性检测、血凝性检测、致病性试验、特异性试验、免疫原性试验以及qPCR和测序鉴定,用病死兔病料组织接种健康易感家兔,成功分离到1株兔出血症病毒。分离株病毒不含细菌、霉菌、支原体;对人"O"型红细泡的血凝效价为1∶1024;分离株病毒能够使健康易感兔在96h内全部死亡兔病毒性出血症;兔出血症病毒阳性血清对分离株病毒具有中和作用;用分离株病毒制备的灭活疫苗具有良好的免疫原性;分离株病毒通过qPCR测序鉴定为RHDV毒株。 相似文献
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旨在了解河南疑似兔出血症病毒2型(RHDV2)的感染情况,并对RHDV2的致病性进行初步分析。本研究采集病死兔的肝组织,利用微量血凝试验、RT-PCR扩增及测序、VP60基因系统进化树分析和动物回归试验进行病原鉴定。微量血凝试验结果显示,组织样本悬液能够凝集人“O”型血红细胞;RT-PCR扩增、测序及序列分析结果显示,检测到RHDV2特异性条带,片段大小为829 bp;系统进化树分析结果发现,分离的病毒与我国四川发现的首例RHDV2毒株SC2020/04的VP60基因相似性高达98.2%;临床病例的剖检显示病死兔胸腺、气管、肺、肝、脾、肾等实质性器官出血较为严重;动物回归试验发现攻毒组家兔死亡率为100%,平均死亡时间为65.8 h,RT-PCR扩增均检测到RHDV2特异性条带。本研究首次在河南兔场检测到RHDV2,为RHDV2的防控提供了科学参考。 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
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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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应用单克隆抗体夹心酶联免疫吸附试验检测兔出血症病毒 总被引：3，自引：0，他引：3
应用建立的单克隆抗体夹心酶联兔疫吸附试验（ＭｃＡｂ－ＥＬＩＳＡ）,检测了人工感染兔出血症病毒（ＲＨＤＶ）ＤＪＲＫ细胞毒、肝毒的兔以及自然感染ＲＨＤＶ的兔的组织样品。结果表明,感染死亡兔的肝、脾、肾、骨髓样品病毒抗原的检出率为１００％,淋巴结和肌肉的检出率分别为９７．５％和７９．５％。ＭｃＡｂ－ＥＬＩＳＡ能检出肌肉中血凝试验不能检出的ＲＨＤＶ抗原。此外,还用ＭｃＡｂ－ＥＬＩＳＡ检测了肝毒人工感染兔血中ＲＨＤＶ的动态,并对１０份兔出血症脏器灭活苗的效价作了滴定。 相似文献
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兔出血症病毒接种幼年兔及乳兔的人工感染试验 总被引：3，自引：0，他引：3
用兔出血症病毒人工接种一月龄与二月龄幼年仔兔，结果证实兔出血症病毒亦能感染并致死幼年仔兔，但感染类型与青成年兔的典型兔出血症不同，主要特征有病程延长，血凝价低或无血凝和黄疸肝炎等等。用兔出血症病毒特异性单克隆抗体建立的ＥＬＩＳＡ试验及反向间接血凝试验自血凝试验阴性的幼年兔各脏器均能检测到兔出血症病毒。人工感染乳兔虽未致临床病症，但扑杀检查见有坏死性肝炎病变和轻度肾小球性肾炎，且自肝肾等血凝检测阴性的病科中，用ＥＬＩＳＡ试验及细胞培养均可检测和分离到病毒。研究结果表明，血凝阴性并不能排除非典型兔病毒性出血症感染。 相似文献
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Marchandeau S Le Gall-Reculé G Bertagnoli S Aubineau J Botti G Lavazza A 《Veterinary research》2005,36(1):53-62
The data were recorded during a Rabbit haemorrhagic disease outbreak that occurred in France in 2001 in a wild population of rabbits that we have been monitoring since 2000. These data suggested the existence of non-protective antibodies due to a putative RHDV-like virus. Twenty-one blood and 22 liver samples were taken from the 26 corpses of recently dead rabbits that were found. RHDV was found in all liver samples. A first screening for RHD antibodies, carried out using an ELISA based on the detection of VP60-RHDV antigen, showed that 20 of the rabbits were seropositive. Moreover, we determined antibody titres for 13 of these 20 seropositive samples. All were > or = 1/400. Such titres normally indicate antibody levels sufficient to confer protection to all known RHDV or RHDV-like strains. For 16 samples, we determined whether these rabbits had died of a chronic or an acute form of the disease, by employing monoclonal antibody (Mabs)--based differential ELISA. All had died of an acute form of RHD. Because the antibodies detected by this VP60-ELISA test are known to appear 5-6 days after infection and since acute RHD generally kills the rabbits 2-3 days after infection, we assumed that the detected antibodies must have been present before the exposure to the virus that killed these rabbits. A second detection of antibodies was made with Mabs that are specific for RHDV. The results were negative, showing that the antibodies detected with the VP60 ELISA test were not specific for RHDV. We sequenced a portion of the VP60 gene of viruses isolated in 17 rabbits. All RHDV isolates were very similar to the RHDV strains commonly isolated in France during this period, suggesting that this viral strain was not a putative variant that is not neutralised by antibodies. Therefore we conclude that the detected antibodies were probably due to a RHDV-like virus that induces the production of detectable but non-protective antibodies. 相似文献
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Teixeira L Marques RM Aguas AP Ferreira PG 《Veterinary immunology and immunopathology》2012,148(3-4):343-347
Rabbit hemorrhagic disease virus (RHDV) is the etiologic agent of rabbit hemorrhagic disease (RHD), an acute lethal infection that kills 90% of adult rabbits due to severe acute liver inflammation. Interestingly, young rabbits are naturally resistant to RHDV infection. Here, we have compared naturally occurring CD4(+)Foxp3(+) regulatory T cells (Tregs) between young and adult rabbits after infection by RHDV. The number and frequency of Tregs was decreased in the spleen of adult rabbits 24h after the RHDV infection; this was in contrast with the unchanged number and frequency of splenic Tregs found in young rabbits after the same infection. Also, serum levels of IL-10 and TGF-β were enhanced in the infected adult rabbits whereas no alteration was observed in infected young rabbits. However, this increase is accompanied by a burst of pro-inflammatory cytokines, but seems not able to prevent the death of the animals with severe acute liver inflammation in few days after infection. Since Tregs downregulate inflammation, we conclude that their decrease may contribute to the natural susceptibility of adult rabbits to RHDV infection. 相似文献
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An internally controlled multiplex real-time RT-PCR using TaqMan probes and external standards for absolute RNA quantification was developed as a new diagnostic tool for the detection of rabbit haemorrhagic disease virus (RHDV). The test revealed a specificity of 100%, an analytical sensitivity of 10 copies/well and a linearity over a range from 10(1) to 10(10) copies. The viral loads in organs, leukocytes, sera and excretions of seropositive, convalescent rabbits which were overcoming an experimental infection with RHDV were determined using the validated assay. As a result, viral RNA was demonstrated and quantified for at least 15 weeks. Thus, a persistence of viral RNA after experimental infection of rabbits could be shown for the first time. In contrast, neither antigen nor infectious virus could be detected by antigen-ELISA, immunohistochemistry or experimental transmission. Therefore, further experiments are necessary to prove that the persistence of RNA is linked with the persistence of infectious virus particles. 相似文献