犬冠状病毒病的诊断与防治措施

正犬冠状病毒病(Canine Coronavirus Disease,CCVD)是由犬冠状病毒(CCV)引起的一种多发性传染病,不同年龄的犬均可感染,该病对幼犬危害严重,发病率和死亡率都较高,是目前对养犬业危害较大的疾病之一。本文系统地对犬冠状病毒病的病原学、流行病学、诊断及防治措施等方面进行综述,以期为犬冠状病毒病的防控提供理论依据。     

犬猫冠状病毒病

由犬猫冠状病毒感染导致的犬猫冠状病毒病对宠物危害较大，不仅影响宠物行业的健康发展，还对人类健康和公共卫生安全构成潜在威胁。本文从病原学、临床症状与病理变化、诊断方法等方面分别对犬冠状病毒病和猫冠状病毒病进行介绍，并对犬猫冠状病毒病的防治进行概述，以期为犬猫养殖者、饲养者及研究人员提供参考。     

犬免疫应激反应的综合防治措施

犬免疫应激反应不但会造成较大的经济损失,还会给犬的免疫预防工作带来负面影响和阻力,本文对犬免疫应激反应的发病原因、临床症状与现实危害进行了阐述,同时结合实际工作提出了综合防治措施。     

犬布鲁氏菌病及其诊治

布鲁氏菌病是危害动物和人类健康的一种人畜共患病。随着生活水平的提高,饲养宠物增多,人类和犬接触越来越密切,犬布鲁氏菌病越来越受到重视。本文针对犬的布鲁氏菌病对其病原学、流行病学、发病机理、检测方法、症状、治疗和免疫预防进行了综述。     

一例犬冠状病毒病的病例报告

犬冠状病毒病是由犬冠状病毒引起的一种临床上以呕吐、腹泻、脱水及易复发为特性的高度接触性传染病。该病对幼犬危害严重,如诊断和治疗不及时,容易导致犬只死亡。就一例犬冠状病毒病的临床诊断及治疗方案进行总结,为提高该病的诊断及治疗水平提供参考。     

兰州市犬冠状病毒病发病情况调查

犬冠状病毒病是由犬冠状病毒引起的以胃肠炎为主的一种高度接触性传染病,对幼犬危害尤其严重,发病率和死亡率都很高,是目前对养犬业危害较大的疾病之一,仅次于犬瘟热、犬细小病毒病和肝炎。采用犬冠状病毒快速检测试剂盒结合临床诊断等方法对兰州市犬冠状病毒病发病情况进行了调查,在总共314份样品中,阳性128份,阳性率为40.76%;而检测的有临床症状的298份样品中,阳性128份,检出率为42.95%;16份临床健康样品,阳性1份,检出率为6.25%,其余全部为阴性。提出了在目前对此病尚无特效的情况下,采取对症活疗和支持疗法,不外乎为不失时机的控制治疗方法。     

犬冠状病毒病的实验室诊断

犬冠状病毒病是由犬冠状病毒 (canine coro-navirus,CCV)引起犬急性胃肠炎的一种高度接触性传染病 ,临床上表现为剧烈的呕吐、腹泻 ,症状消失后 2～ 3周 ,还可能复发 ,是当前对养犬业危害较大的疫病之一 [1] 。由于犬冠状病毒的流行病学特点、临床症状和病理剖检均缺乏特征性变化 ,在血液学和生物化学方面也没有特定指标 ,故对于该病的确诊往往必须通过实验室诊断。近年来 ,国内外在犬冠状病毒病诊断方面的研究十分活跃 ,尤其是在分子病毒学诊断方面取得了可喜进展 ,现就这些方法综述如下。1　电子显微镜和免疫电镜观察取粪便悬液作负染后…     

辽宁省警犬犬瘟热犬细小病毒和犬冠状病毒感染情况调查

犬传染病是危害养犬业健康发展的主要疾病,来自警犬方面的统计资料证实,全国每年免疫警犬的发病率为14％,其中传染病占60％以上,因传染病而死亡的犬超过死亡犬总数的80％。从近几年的临床实践和国外的有关报道来看,危害严重的犬病毒病主要有犬瘟热、犬细小病毒感染、犬传染性肝炎、犬冠状病毒病、犬副流感及狂犬病等6种疾病,     

中西医结合治疗犬冠状病毒病

犬冠状病毒病是由犬冠状病毒引起犬急性胃肠炎的一种高度接触性传染病。近几年来,我区养犬业发展迅速,犬冠状病毒病日益流行,给养犬业带来很大的危害,犬不分品种、年龄、性别都可感染,若不及时治疗可引起死亡。笔者自2011年以来,先后收治多例冠状病毒感染的病例,并摸索出较为理想的综合治疗方法及预防措施,治愈率达98％。     

浅析犬瘟热、犬细小病毒病的诊治

犬病毒性传染病包括犬瘟热、犬细小病毒病、犬传染性肝炎、狂犬病、犬冠状病毒病、犬疱疹病毒病、犬轮状病毒病、犬副流感、伪狂犬病等.下面笔者主要对临床中最为常见,且发病率、死亡率高,对犬类危害严重的犬瘟热、犬细小病毒病这两种病毒性传染病的诊断及防治作一介绍.     

Isolation and identification of a canine coronavirus strain from giant pandas (Ailuropoda melanoleuca)

Two giant pandas (Ailuropoda melanoleuca) died of unknown causes in a Chinese zoo. The clinical disease profile suggested that the pandas may have suffered a viral infection. Therefore, a series of detection including virus isolation, electron microscopy, cytobiological assay, serum neutralization and RT-PCR were used to identify the virus. It was determined that the isolated virus was a canine coronavirus (CCV), on the basis of coronavirus, neutralization by canine anti-CCV serum, and 84.3% to 100% amino acid sequence similarity with CCV. The results suggest that the affected pandas had been infected with CCV.     

犬冠状病毒胶体金检测卡制备与评价

为快速检测犬冠状病毒,研究利用胶体金免疫层析技术制备和评价犬冠状病毒抗原检测卡。试验通过将胶体金标记的CCV Ab1和鼠IgG涂布于聚酯纤维膜上作为标记垫,生物素耦联的CCV Ab2处理在样品垫上,链霉亲和素和羊抗鼠IgG包被于NC膜上分别为检测线和质控线,与吸水纸和PVC底板制备犬冠状病毒抗原胶体金检测卡,并对其评价。结果:该检测卡不与其他病毒发生交叉反应,可检测浓度低至2 ng/mL的样本,24月是稳定的。结论:该测试板具有良好的特异性、灵敏度、重复性和稳定性,并且与RT-qPCR的结果较符合。因此,该检测卡能快速、准确地检测犬冠状病毒,为动物临床诊断和治疗提供参考。     

基因芯片检测5种动物冠状病毒的初步研究

采用蔗糖密度梯度离心,纯化浓缩犬冠状病毒(CCV)、猫冠状病毒(FCV)、猫传染性腹膜炎病毒(FIPV)、猪传染性胃肠炎病毒(TGEV)、猪呼吸道冠状病毒(PRCV)的细胞培养物,分别设计7,17,11,10和4对引物,构建了49个基因片段的克隆。煮沸裂解法制备质粒DNA,回收PCR扩增产物,点制冠状病毒基因芯片。抽提病毒总RNA,利用Cy3-dCTP随机渗入反转录PCR标记,与芯片进行杂交检测,淘汰交叉的克隆片段。结果表明:克隆CCV1,CCV2,CCV5和CCV7可特异诊断CCV,克隆FCV6,FCV7,FCV8和FCV9可特异诊断FCV,克隆FIPV2,FIPV7,FIPV8和FIPV9可特异诊断FIPV,克隆PRCV1,PRCV2和PRCV3可特异诊断PRCV,克隆TGEV3,TGEV4,TGEV5和TGEV6可特异诊断TGEV。将这些特异克隆扩增片段重新点制基因芯片,与病毒PCR产物杂交,未发现交叉现象。基因芯片检测比传统PCR敏感1000倍,可有效应用于这5种动物冠状病毒的检测与区分。     

Canine coronavirus-associated puppy mortality without evidence of concurrent canine parvovirus infection.

This report presents 2 cases in which puppy fatalities were associated with canine coronavirus (CCV), but no evidence of concurrent canine parvovirus (CPV-2) disease was observed. Case 1 involved a 7-week-old, male short-haired Chihuahua, which had become lethargic 24 hours after purchase from a pet store. Within 72 hours, the puppy began to vomit, had diarrhea, and was admitted to the veterinary clinic, where it was placed on IV fluids. The parvovirus Cite test was negative. The puppy died within 12 hours of admission and was submitted for diagnostic workup. Gross pathology revealed an enteritis suggestive of CPV-2. Histopathology on intestines showed scattered dilated crypts with necrotic cellular debris and neutrophils. There was moderate depletion and necrosis of lymphoid follicles. Electron microscopy (EM) on intestinal contents was positive for coronavirus and negative for parvovirus. Immunohistochemistry (IHC) on gut sections was positive for CCV and negative for CPV-2. Case 2 was an 8-week-old, male Shih Tzu, which was admitted to the veterinary clinic exhibiting symptoms of severe gastroenteritis with abdominal pain. The referring veterinarian euthanized the puppy, and the entire body was submitted for diagnostic evaluation. Necropsy revealed a severe ileo-cecal intussusception and segmental necrotic enteritis of the small intestine. Electron microscopy of the intestinal contents was positive for coronavirus and negative for parvovirus. Immunohistochemistry on sections of affected gut were positive for CCV and negative for CPV-2. These cases emphasize the importance of pursuing a diagnosis of CCV in young puppies when CPV-2 disease has been ruled out by IHC.     

Induction and enhancement of feline infectious peritonitis by canine coronavirus.

Preexisting antibody to feline infectious peritonitis virus (FIPV) causes acceleration and enhancement of disease on subsequent infection of cats with FIPV. Other workers have shown that canine coronavirus (CCV) can infect cats subclinically, but have found no evidence of enhancement of, or protection against, subsequent FIPV infection. With various isolates of CCV, we determined that 1 strain of CCV can induce transient mild diarrhea in cats and, furthermore, that previous infection with CCV causes acceleration and enhancement of subsequent infection with FIPV. In addition, sequential inoculation of cats with another strain of CCV caused lesions indistinguishable from those of FIP, without exposure at any time to FIPV.     

Differentiation of canine coronavirus and porcine transmissible gastroenteritis virus by neutralization with canine,porcine and feline sera

Monospecific antisera were prepared in rabbits against canine coronavirus (CCV) and transmissible gastroenteritis virus of pigs (TGEV), and in 24 pigs and 3 cats against TGEV alone. Neutralizing antibody titres were higher for the immunizing than the heterologous virus, although cross-neutralization usually was detected. This confirmed that CCV and TGEV are distinct, but antigenically related coronaviruses. In sera from 41 dogs, CCV-neutralizing titres were on average 2.7 fold higher than TGEV-neutralizing titres, suggesting that CCV was the causal agent. Sera from 29 cats in colonies with feline infectious peritonitis (FIP) and known to contain TGEV-neutralizing antibody, were found to have titres 12.3 fold higher against CCV. The FIP virus (FIPV) is probably more closely related to CCV than TGEV as judged by antigens involved in virus neutralization.Antisera to two isolates of bovine coronavirus, three isolates of haemagglutinating encephalomyelitis virus, seven strains of avian infectious bronchitis virus and the 229E strain of human coronavirus all failed to neutralize CCV and TGEV. Thus CCV, TGEV and probably FIPV fall into a group of antigenically related agents, separable from other members of the family Coronaviridae, by both virus neutralization and immunofluorescence tests.     

Enzyme-linked immunosorbent assay for the detection of canine coronavirus and its antibody in dogs.

Two methods of enzyme-linked immunosorbent assay (ELISA) were developed for the diagnosis of canine coronavirus (CCV) infection in dogs. One ELISA, in which CCV-infected CRFK cell lysate is used as antigen, is for the detection and titration of antibody against CCV, and the other ELISA uses the double antibody sandwich method for the detection of CCV antigen. The first ELISA procedure demonstrated antibody responses in dogs inoculated with CCV, as did the virus neutralization test; the second ELISA detected specific CCV antigen in feces and organ homogenates of inoculated dogs.     

犬冠状病毒N蛋白的原核表达及其多克隆抗体的制备

本研究旨在克隆犬冠状病毒（canine coronavirus,CCV）N基因,体外表达N蛋白,并制备抗该蛋白质的多克隆抗体,用于CCV的诊断及其抗原的检测。参考GenBank中CCV的N基因序列（登录号：KY063618.2）,选择CCV流行毒株的N基因,通过对该基因密码子进行优化和基因合成,最后选择一段有效基因构建重组表达质粒pET-B2M-N,将成功构建的重组质粒转化大肠杆菌DH5α感受态细胞,挑取阳性克隆提取质粒,转化大肠杆菌BL21（DE3）感受态细胞,通过0.5 mmol/L IPTG 30 ℃进行诱导表达。结果表明,优化诱导条件后成功表达出大小约为49 ku的重组蛋白。将重组蛋白与弗氏佐剂按一定比例混合,每隔2周免疫G767、G768两只日本大耳白兔数次,用间接ELISA检测G768抗体效价可达1∶512 000,选用G768抗体进行抗体纯化,纯化后浓度可达10 mg/mL,用间接ELISA、Western blotting和间接免疫荧光试验对N蛋白纯化后制备的兔多克隆抗体进行检测分析,表明表达的重组N蛋白免疫原性良好,制备的多克隆抗体具有良好的反应原性。本研究为犬冠状病毒抗原抗体检测以及靶标CCV诊断试剂盒的建立奠定了一定的基础。     

ELISA法检测犬腹泻粪样中的犬冠状病毒

用ＦＥ细胞增殖犬冠状病毒（ＣＣＶ）参考株，分别免疫家兔和ＢＡＬＢ／ｃ小鼠制备ＣＣＶ多抗和单抗，建立了夹心ＥＬＩＳＡ及Ｄｏｔ－ＥＬＩＳＡ诊断方法。在检测的８４例犬腹泻粪样中，多抗、单抗夹心法显示ＣＣＶ阳性１６例，Ｄｏｔ－ＥＬＩＳＡ阳性１３例，后１３例包括在前１６例中。从８４例腹泻犬粪样中随机取３８例作ＣＣＶ、犬细小病毒（ＣＰＶ）双项检测，ＣＣＶ阳性１６例，ＣＰＶ阳性６例，ＣＣＶ、ＣＰＶ混合感染４例。结果显示，在南京地区流行的犬腹泻中，ＣＣＶ感染比例有超过ＣＰＶ的趋势。