鸭瘟病毒PCR-DHPLC检测方法的建立

为了快速、准确诊断和防制鸭瘟疫病,试将PCR技术和变性高效液相色谱技术(DHPLC)相结合建立新的诊断方法,对鸭瘟进行诊断。结果表明:PCR-DHPLC技术是一种快速、高敏感性、高特异性的检测方法。     

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山东 陈洪：鸭霍乱与鸭瘟诊断方法有哪些？ 袁圆：一、手摸诊断。患鸭霍乱病的病鸭，死亡之前常摇头，死亡时，嘴、鼻流血水，嗉囊里充满饲料，手摸嗉囊感觉较硬；而患鸭瘟的病鸭，流涕流眼泪，死亡时眼睛充血，嗉囊空空无食物，手摸嗉囊较松软。     

袁圆在线

山东陈洪：鸭霍乱与鸭瘟诊断方法有哪些?袁圆：一、手摸诊断。患鸭霍乱病的病鸭,死亡之前常摇头,死亡时,嘴、鼻流血水,嗉囊里充满饲料,手摸嗉囊感觉较硬；而患鸭瘟的病鸭,流涕流眼泪,死亡时眼睛充血,嗉囊空空无食物,手摸嗉囊较松软。二、药物与病程诊断。鸭霍乱一般是零星发生,突然死亡,尤其是正在产蛋的母鸭较为多见,鸭霍乱病还能传染给鸡、猪等,而鸭瘟流行范围较广,不能传染给鸡和猪,一般多在发病后5天左右死亡。鸭霍乱可用磺胺类药物或抗菌素治疗,但对治疗鸭瘟却无效。     

新型鸭瘟的诊断和防治

新型鸭瘟(new duck plague)是一种由新型疱疹病毒引起的传染病.该病同鸭瘟的临床症状和病理变化都极为相似,使用已有的鸭瘟疫苗、高免血清、高免卵黄抗体及药物均不能有效防治该病,给我国养鸭业造成了严重的经济损失.对山东省某养鸭场发病症状比较明显的病鸭采取临床剖检、病理组织学观察、病毒的分离鉴定以及血清学检查等诊断措施,确诊该鸭场病鸭感染的疾病是新型鸭瘟.详细介绍了该次新型鸭瘟的诊断过程,旨在为生产中新型鸭瘟的诊断提供参考.     

一起鸭瘟的诊制

鸭瘟是由鸭瘟病毒引起的一种急性败血性传染病.本市一蛋鸭场饲养4 000多只蛋鸭,145日龄开始发病,1周内死了40多只,诊断为鸭瘟.     

鸭瘟的综合防治

鸭瘟是由鸭瘟病毒引起的鸭的一种急性、热性、败血性传染病。临床特征为高热,两腿发软无力,下痢,口渴,流泪和部分病鸭头颈部肿大,俗称大头瘟。本病传播迅速,发病率和死亡率都高。本文就鸭瘟病毒的研究历史、病原学、流行病学、临床症状、病理变化、诊断、治疗预防等研究进行了一次综合整理,希望为鸭瘟的研究提供一些理论上的帮助。     

鸭瘟的诊断与防制

<正>鸭瘟是鸭、鹅的急性、热性、败血性传染病,该病传播迅速,发病率和死亡率都很高,是一种严重危害养鸭业的疾病。本文就鸭瘟的流行病学、临床诊断及防制方法等作简要的概述,以为广大     

鸭鹅常见传染病诊断与防治

笔者介绍9种鸭鹅常见传染病鸭瘟、鸭病毒性肝炎、雏番鸭细小病毒病、禽霍乱、传染性浆膜炎、鸭流感、小鹅瘟、副黏病毒病、鹅出血性坏死性肝炎的诊断与防治方法.     

一起樱桃谷鸭鸭瘟的诊断报告

鸭瘟俗称大头瘟,是由鸭瘟病毒引起的一种急性败血性传染病.2009年9月某养鸭户从外地购得樱桃谷苗鸭1000余只进行饲养,不到1个月就陆续发病全部死亡,经流行病学调查、病理剖检、实验室诊断,确诊为鸭瘟,现将有关情况报告如下:     

鸭急性传染病的鉴别诊断

近几年来,急性鸭病的发生不但给广大畜牧兽医工作者临床诊断带来了困难,而且还给养殖场造成不必要的经济损失。现将急性死亡的鸭传染病的鉴别诊断总结如下。一、外观鉴别要点1.鸭霍乱、鸭副伤寒使鸡鸭都发病。2.鸭瘟、鸭病毒性肝炎仅发病于鸭。鸭瘟的成鸭发病多于幼鸭,鸭病毒性肝炎的雏鸭发病多于成鸭。     

Identification of duck plague virus by polymerase chain reaction

A polymerase chain reaction (PCR) assay was developed for detecting duck plague virus. A 765-bp EcoRI fragment cloned from the genome of the duck plague vaccine (DP-VAC) virus was sequenced for PCR primer development. The fragment sequence was found by GenBank alignment searches to be similar to the 3' ends of an undefined open reading frame and the gene for DNA polymerase protein in other herpesviruses. Three of four primers sets were found to be specific for the DP-VAC virus and 100% (7/7) of field isolates but did not amplify DNA from inclusion body disease of cranes virus. The specificity of one primer set was tested with genome templates from other avian herpesviruses, including those from a golden eagle, bald eagle, great horned owl, snowy owl, peregrine falcon, prairie falcon, pigeon, psittacine, and chicken (infectious laryngotracheitis), but amplicons were not produced. Hence, this PCR test is highly specific for duck plague virus DNA. Two primer sets were able to detect 1 fg of DNA from the duck plague vaccine strain, equivalent to five genome copies. In addition, the ratio of tissue culture infectious doses to genome copies of duck plague vaccine virus from infected duck embryo cells was determined to be 1:100, making the PCR assay 20 times more sensitive than tissue culture for detecting duck plague virus. The speed, sensitivity, and specificity of this PCR provide a greatly improved diagnostic and research tool for studying the epizootiology of duck plague.     

A Plaque Assay for Duck Plague Virus

A plaque assay for duck plague virus was developed for a chicken embryo-adapted virus and a duck lethal virus and used to determine the identity of these viruses. Using the plaque inhibition neutralization test, duck plague virus was differentiated from Newcastle disease, fowl plague, and duck hepatitis viruses. The plaque morphology is described.     

Diagnosis of duck plague in waterfowl by polymerase chain reaction

A recently developed polymerase chain reaction (PCR) assay was used for diagnosis of duck plague in waterfowl tissues from past and current cases of waterfowl mortality and to identify duck plague virus in combined cloacal/oral-pharyngeal swab samples from healthy mallards (Anas platyrhynchos) after a disease outbreak. The PCR was able to detect viral DNA from all the individual or pooled tissues assayed from 10 waterfowl, including liver and spleen samples from three Muscovy ducks (Cairina moschata domesticus) that did not yield virus isolates. The strong staining intensity of the PCR products from the waterfowl tissues indicated that large amounts of virus were present, even when virus was not isolated. Duck plague DNA was also detected in a cloacal swab sample from a wood duck (Aix sponsa) carcass submitted for diagnosis. The PCR assay identified duck plague DNA in 13 swab samples that produced virus isolates from carrier mallards sampled in 1981 after a duck plague die-off. The duck plague PCR clearly demonstrated the ability to quickly diagnose duck plague in suspect mortality cases and to detect virus shed by carrier waterfowl.     

Superinfection in ducks persistently infected with duck plague virus

Superinfections with homologous or heterologous strains of duck plague virus resulted in the deaths of birds persistently infected with duck plague virus. Not all birds that were superinfected died. Protection against mortality depended on the route of exposure, strain of the initial duck plague virus, and strain of the superinfecting virus.     

番鸭细小病毒与鸭圆环病毒二重PCR方法的建立

根据基因库中鸭圆环病毒和番鸭细小病毒的基因序列,分别设计了两对特异性引物,通过对二重PCR扩增条件的优化,研究建立了可同时鉴别检测鸭圆环病毒和番鸭细小病毒的二重PCR方法。用该方法对同一样品中鸭圆环病毒和番鸭细小病毒的模板进行PCR扩增,结果均得到了与实验设计相符的351bp（鸭圆环病毒）和474bp（番鸭细小病毒）的扩增条带,而对鸭Ⅰ型肝炎病毒、鹅细小病毒、鸭副黏病毒、鸭瘟病毒和禽流感病毒等病原体的检测全为阴性。敏感性测定结果表明：该二重PCR技术最低能检出100fg的鸭圆环病毒和番鸭细小病毒DNA模板。研究建立的鸭圆环病毒和番鸭细小病毒的二重PCR方法,具有快速、敏感、特异、定量和重复性好等优点,可用于临床上鸭圆环病毒和番鸭细小病毒感染的检测。     

Outbreak of duck plague (duck herpesvirus enteritis) in numerous species of captive ducks and geese in temporal conjunction with enforced biosecurity (in-house keeping) due to the threat of avian influenza A virus of the subtype Asia H5N1

The continuing westward spread of avian influenza A virus of the subtype H5N1 in free-living and domestic birds forced the European Union and the German federal government to enhance all biosecurity measures including in-house keeping of all captive birds from October 20 to December 15, 2005. Movement of captive ducks and geese of many different species from a free-range system to tight enclosures and maintenance for prolonged times in such overcrowded sheds resulted in pronounced disturbance of natural behaviour, interruption of mating and breeding activities and possibly additional stress. Under these conditions the birds developed signs of severe disease and enhanced mortality twentyfour days later. A total of 17 out of 124 (14%) adult birds and 149 out of 184 year-old birds (81 %) died during the outbreak. A herpesvirus was isolated from many organs of succumbed ducks and geese that was identified as a duck plague herpesvirus by cross neutralization test using known antisera against duck plague virus. The published host range of duck plague comprises 34 species within the order Anseriformes. We report here on additional 14 species of this order that were found to be susceptible to duck plague virus. The exact source of the herpesvirus could not identified. However, low antibody titres in some ducks at day of vaccination indicate that at least some of the birds were latently infected with a duck plague herpesvirus. The remaining healthy appearing birds were subcutaneously vaccinated with a modified live duck plague vaccine (Intervet, Boxmeer, NL) that stopped losses and resulted in seroconversion in most of the vaccinated birds.     

鸭新城疫病毒和鸭圆环病毒二重PCR检测方法的建立

本研究根据GenBank中鸭新城疫病毒(NDV)的F基因和鸭圆环病毒(DuCV)的V1/rep基因的保守序列,各设计一对特异性引物,并对二重PCR的扩增条件进行优化,建立了鸭NDV和DuCV的二重PCR检测方法。对混合样品进行扩增,得到2条大小为493bp(鸭NDV)和218bp(DuCV)的特异性条带,与预扩增片段相符。而对番鸭细小病毒、鸭瘟病毒、鸭肝炎病毒、鸭源小鹅瘟病毒、鸭H9亚型流感病毒、鸭疫里氏杆菌、大肠杆菌、禽多杀性巴氏杆菌等病原检测,结果为阴性。该方法的敏感性试验表明,鸭NDV的核酸最小量为40fg,DuCV为20fg。     

The occurrence of virus enteritis (duck plague) in wild birds]

296 samples from wild birds of 15 species were incorporated into long term exploration of duck plague on ducks in farms. By using virological and serological standard methods 9 virus carriers and 20 serum samples showing positive antibody titers could be detected. The epidemiology as well as the relation of the incidence of duck plague in wild birds and farm poultry is discussed.     

番鸭三种细小病毒野毒株试验感染雏鸭的临床症状及解剖病理变化差异研究

番鸭"三周病"、细小病毒型"白点病"和小鹅瘟是雏番鸭常见的三种细小病毒病,对番鸭养殖业危害较为严重。本文对从广东惠州及周边地区病死番鸭中分离鉴定的1株番鸭细小病毒型"白点病"病毒、1株小鹅瘟病毒和1株番鸭"三周病"病毒分别感染健康雏番鸭,将发病的临床症状、病理学变化进行对比,从而为基层兽医工作者提供三种番鸭细小病毒病临床鉴别、初步诊断更加直观的依据。     

鸭瘟活疫苗外源病毒检验方法（鸡检查法）的修订

对二〇〇五年版《中华人民共和国兽药典》中鸭瘟活疫苗外源病毒检验的鸡检查法进行了修订,确定选用9-12周龄SPF鸡,在原药典规定方法的基础上增加低剂量基础免疫程序,并用该外源病毒检验方法对5批鸭瘟活疫苗进行了验证,结果表明修订后的鸭瘟活疫苗外源病毒检验方法可靠,可操作性强。