静水压技术新城疫病毒灭活工艺制备油乳剂疫苗的研究

病毒在静水压力50、200、250、300、350MPa灭活时间分别为60、50、30、20、20min,结果表明静水压力处理病毒颗粒体积缩小,免疫原性没有破坏;压力越高病毒失活越快,低压力长时间与高压力短时间灭活病毒的效果相同,以传统灭活制备的疫苗为对照,研究静水压技术新城疫病毒灭活工艺制备油乳剂疫苗的免疫原性。     

犬瘟热病毒高压灭活疫苗的制备与应用

本试验用静水压高压灭活的方法对ＣＤＶ进行灭活,并加入蜂胶佐剂制备犬瘟热病毒高压灭活疫苗。将此灭活疫苗免疫接种ＣＤＶ易感犬,同时以用福尔马林灭活的犬瘟热病毒灭活疫功为对照,检测结果表明：犬瘟热病毒高压灭活疫苗安全、可靠,其诱导试验犬所产生中和抗体的能力明显高于该病毒的福尔马林灭活苗,临床应用证明该疫苗对犬的保护率达８９％。     

犬瘟热组织脏器灭活苗的制备与应用

应用PCR方法对多个毛皮动物养殖场收集了大量的疑似犬瘟热的病料进行检测,筛选的阳性病料用甲醛按常规方法制备组织脏器灭活苗,并进行无菌检验、安全试验和临床应用实验.结果显示制备的组织脏器灭活苗不含有任何细菌,对幼狐免疫后无明显的不良反应,临床紧急接种应用实验显示该疫苗有效率高达83.3%.表明该组织脏器灭活苗安全有效,可应用于毛皮动物犬瘟热的紧急接种.     

犬瘟热病毒R／20-8株核衣壳蛋白基因的克隆和表达

应用RT-PCR技术扩增出犬瘟热病毒（CDV）核衣壳（N）蛋白基因的高度保守序列，将其克隆至质粒pMD18-T中，获得了重组质粒pMD18-T-N。将N基因的目的片段克隆到表达载体pGEX-4T-1中谷胱甘肽转移酶（GST）基因的下游，并将该重组质粒转化大肠杆菌BL21株，经IPTG诱导，N基因融合蛋白获得了高效表达。SDS-PAGE电泳和Western—blot分析结果显示，表达产物的分子质量为55ku，与CDV标准阳性血清呈阳性反应。表明，大肠杆菌表达的CDVN蛋白在免疫原性上与天然N蛋白具有一定的相似性，可作为诊断用抗原。     

犬瘟热病毒与犬瘟热的预防免疫

犬瘟热(Canine Distemper,CD)是由犬瘟热病毒(Canine Distemper.Virus,CDV)感染引起的急性、高度接触性传染病.它常引起大批犬、貂、狐等动物发病,病死率达30%～80%.是当前对我国养犬业、毛皮动物养殖业和野生动物保护业危害最大的疫病之一.因此提高对犬瘟热的预防和控制也是当前值得关注的问题.     

犬瘟热病毒的分子生物学研究进展

犬瘟热病毒的分子生物学研究进展耿志贤田克恭（军事医学科学院实验动物中心，北京１００８５０）李钟铎（军事医学科学院微生物学流行病学研究所）犬瘟热病毒（ｃａｎｉｎｅｄｉｓｔｅｍｐｅｒｖｉｒｕｓ，ＣＤＶ）是犬瘟热病的病原。自然条件下ＣＤＶ感染食肉目中犬科、...     

犬瘟热病毒的检测技术

从免疫学和分子生物学两方面对犬瘟热病毒检测技术进行了综述 ,并探讨了今后的研究方向。     

犬瘟热病毒H基因的序列分析与表达

以临床疑似犬瘟热病犬外周血总RNA为模板,采用RT-PCR方法扩增犬瘟热病毒(CDV)血凝素(H)基因。序列测定和分析表明该病料犬瘟热病毒H基因序列与国内其他地区分离到的毒株同源性较高(94.6%~99.2%),而与Onderstepoort株、Convac株等疫苗毒株的同源性较低(90.4%~91.2%)。分子遗传进化分析显示所有毒株可分为7个大的分支,且各分支间具有一定的地域性,其中待检病料中CDV与大多数中国分离株一样处于Ⅰ型。进一步以H全基因为模板,截短表达其3′端816 bp、459 bp两个片段,并克隆入原核表达载体pET30 a进行表达。结果显示它们分别能表达大小约为36.4 ku和22.2 ku的融合蛋白。免疫转印表明该纯化蛋白均可与犬瘟热病毒抗血清发生阳性反应,说明重组蛋白具备抗原性,可用于进一步的血清学研究。     

Immunoperoxidase study of canine distemper virus pneumonia

Forty-two cases of canine pneumonia were examined for the presence of canine distemper virus. For that purpose canine distemper virus inclusion bodies were located. The histopathological lesions were related to the presence of canine distemper antigen, as demonstrated with an immunoperoxidase technique. This technique was more sensitive for detecting canine distemper infection in lung tissue than was the study of inclusion bodies. Attention was also paid to combined infection with canine adenovirus and Bordetella bronchiseptica.     

Neurological manifestations of canine distemper virus infection

Canine distemper virus causes a multi systemic disease in dogs often with severe neurological signs. These signs are the result of viral replication in neurons and glial cells leading to grey matter lesions and demyelination. Inflammation leads to further destruction of the tissue. As extraneural signs are often lacking and only one localisation may be found on neurological examination, distemper may be difficult to diagnose. Myoclonus is almost pathognomonic for this disease but occurs in less than half of the cases. The inflammation of the central nervous system that occurs during the chronic stage of the disease can be detected on examination of the cerebrospinal fluid, in particular by determination of the IgG index. Viral antigen can be demonstrated in cerebrospinal fluid cells by fluorescent antibody techniques. The prognosis of nervous distemper is generally poor although dogs can recover from this disease. Treatment is largely supportive and symptomatic. The importance of regular vaccination is stressed.     

犬瘟热病毒感染的研究现状

犬瘟热（Canine distemper）是由犬瘟热病毒（Canine distemper virus,CDV）引起犬的一种急性、高度接触性传染病。早期表现以双相热、急性鼻卡他、卡他性肺炎和胃肠炎为特征,后期出现神经症状,少数病犬出现鼻和足垫过度角化现象。CDV感染是犬科动物最常见的、最主要的病毒性疾病之一,发病率高、治愈率低,是食肉动物病毒病的典型代表。笔者就近年来我国CDV感染的研究现状简述如下。     

Polioencephalomalacia associated with canine distemper virus infection

Eight dogs with severe neurologic signs, including seizures, had polioencephalomalacia of the pyriform cortex, Ammon's horn and deep structures in the temporal lobe. The polioencephalomalacia was considered to be a consequence of canine distemper virus infection based on clinical signs, typical inclusions, the demonstration of viral antigens in the lesions and of characteristic paramyxovirus nucleocapsids by electron microscopy. Little evidence for neuronal destruction by direct viral activity was found. Selective nerve cell necrosis was attributed to ischemia (vascular lesions and seizure induced consumptive anoxia) and immune mechanisms. The selective involvement of the rhinencephalic structures was thought to be related to the mode of entry and spread of the virus.