用ELISA法检测犬细小病毒抗体

本文采用吸附豚鼠犬细小病毒（ＣＰＶ）多克隆抗体的微量板，进行间接酶免疫测定（ＥＬＩＳＡ）检测犬血清中的ＣＰＶ抗体。结果，ＥＬＩＳＡ抗体效价比血凝抑制抗体（ＨＩ）效价高，与ＨＩ效价的相关系数ｒ＝０．７８。     

对流免疫电泳检测犬细小病毒免疫复合物中抗体的研究

通过对流免疫电泳的条件优选,实现了解离后犬细小病毒复合物中抗原抗体的分,达到了检测免疫复合物中抗体的目的。在应用检测试验中,通过与酶档ＳＰＡ染色法、血凝抑制试验及常规对流免疫电泳法比较,证实了该种检测方法的可靠性、敏感性和实用价值。     

几个猫细小病毒毒株间抗原性和基因性的比较

用病毒ＤＮＡ限定酶及抗猫泛白细胞减少症病毒（ＦＰＬＶ）单克隆抗体对源于猫、水貂及犬的１４株猫细小病毒（ＦＰＶ）进行抗原和基因（差异）的比较测定。且在抗原性及基因特性方面与日本的ＦＰＬＶ及水貂肠炎病毒（ＭＥＶ）毒株进行比较。最近从中国东北部分离出来的几株ＭＥＶ和狗细小病毒（ＣＰＶ，“新”抗原型）具更多的相同之处。由于抗原及基因特性所决定我们还是把一份从普通猫粪便中分离出的一个毒株看作ＣＰＶ。用限定酶（ＨｉｎｆＩ）消化显示。早期分离到的一株“新”抗原型ＣＰＶ和分离到的“旧”抗原型ＣＰＶ毒株具有相似的消化片断。包括上述提到的这些特性的测定结果显示．在ＦＰＶ的不同亚种间存在抗原的多相性和基因的多型性。     

应用间接ELISA检测犬冠状病毒抗体

为了建立一种快速、敏感、特异的检测犬冠状病毒（ＣＣＶ）的方法,利用感染ＣＣＶ的犬肾传代细胞包被酶联反应板,以辣根过氧化物酶标记的金黄色葡萄球菌Ａ蛋白（ＨＲＰ－ＳＰＡ）作为第２抗体,建立了检测ＣＣＶ抗体的间接ＥＬＩＳＡ方法。抗原的包被量为每孔３×１０４个染毒细胞,酶标抗体的工作浓度为１∶４０。试验发现,包被的病毒抗原经甲醇固定３０ｍｉｎ后可以提高试验的敏感性,减少病毒抗原的使用量。与中和试验比较证实,２种抗体检测方法的测定结果呈正相关     

犬瘟热、犬细小病毒特异转移因子的研究

以免疫犬脾为原料，应用静置透析法，制备了犬瘟热、犬细小病毒特异转移因子（ＣＰＶ，ＣＤＶ—ＳＰＴＦ），并进行了理化性质及生物学特性检测。结果表明：ＣＰＶ；ＣＤＶ—ＳＰＴＦ无菌、无热原、无过敏性、不含大分子蛋白质、能增加受体Ｔ细胞比值、特异性提高ＣＰＶ、ＣＤＶ抗体水平，和疫苗联用时有协同作用，增强疫苗效果，具有对细胞免疫和体液免疫的双重调节作用。     

犬细小病毒病的诊疗

犬细小病毒病的诊疗张智勇（河南职业技术师范学院畜牧兽医系，辉县４５３６００）犬细小病毒病是由犬细小病毒（ＣＰＶ）引起的一种急性传染病，又称犬传染性出血性肠炎。１９９３～１９９５年，笔者利用我校动物医院接诊的１２６２例病例，开展了ＣＰＶ病诊疗方法的研究...     

北京地区腹泻犬细小病毒感染情况调查

在校动物医院用试剂盒对１３６例门诊腹泻犬粪便进行犬细小病毒（ＣＰＶ）检验，其中阳性７２例（阳性率５２．９％）。阳性病例中，公犬占５２．４％，母犬占５１．６％，因此ＣＰＶ感染与犬性别无关。ＣＰＶ阳性率，以２～４月龄大为最高（６０．３％），５～６月龄次之（１７．６％）。７月龄以后较少。未注射或未按程序注射ＣＰＶ疫苗犬，ＣＰＶ检验阳性率高，即使按程序接种ＣＰＶ疫苗的犬，也仍有感染ＣＰＶ的，其原因有待研究．     

北京地区腹泻犬细小病毒感染情况调查

在校动物医院用试剂盒对１３６例门诊腹泻犬粪便进行犬细小病毒（ＣＰＶ）检验，其中阳性７２例（阳性率５２．９％）。阳性病例中，公犬占５２．４％，母犬占５１．６％，因此ＣＰＶ感染与犬性别无关。ＣＰＶ阳性率，以２－４月龄犬为最高（６０．３％），５－６月龄次之（１７．６％），７月龄以后较少。未注射或未按程序注射ＣＰＶ疫苗犬，ＣＰＶ检验阳性率高，即使按程序接种ＣＰＶ疫苗的犬，也仍有感染ＣＰＶ的，其原因有待研究     

犬细小病毒性肠炎

犬细小病毒性肠炎是由犬细小病毒 (ＣＰＶ)引起的以剧烈呕吐和腹泻及白细胞明显减少为主要临床特征的犬细小病毒性肠炎。本病作为一种急性、热性传染病 ,危害极其严重。1　病原与流行情况该病的病原为犬细小病毒 (ＣＰＶ) ,属犬细小病毒科细小病毒属。病毒可在犬、狼、狐狸等动     

动物细胞系的染色体组型分析和致癌／致瘤性

为确定供生物制品和生物工程产品生产所用动物传代细胞系有无致癌／致瘤性,对研制生产犬、貉、狐用五联病毒,即犬狂犬病病毒（ＲＶ）、犬瘟热病毒（ＣＤＶ）、犬细小病毒（ＣＤＶ）、犬腺病毒２型（ＣＡＶ２）和犬副流感病毒（ＣＰＩＶ）的活疫苗以及猫、狮、虎、豹用三联病毒,即泛白细胞减少症病毒（ＦＰＬＶ）、猫疱疹病毒１型（ＦＨＶ１）和猫杯状病毒（ＦＣＶ）的活疫苗,所用猫肾细胞系（Ｆ８１,ＣＲＦＫ）、犬肾细胞系（ＭＤＣＫ）、非洲绿猴肾细胞系（Ｖｅｒｏ,Ｖｅｒｏ２）、恒河猴肾细胞系（ＭＡ１０４）和叙利亚金…     

Establishment and Application of Colloidal Gold Immune Chromatography Test Method for Detection of Canine Parvovirus Hemagglutination Inhibition Antibody Titer

The study was aimed to use colloidal gold immune chromatography technology to establish a rapid method for detection of canine serum canine parvovirus (CPV) hemagglutination inhibition (HI) titer and CPV vaccine immunization effect assessment.Double antibody sandwich method and monoclonal antibodies of anti-CPV hemagglutination antigen were used to prepare CPV antigen test strip.Canine serum with different proportion respectively was mixed with quantitative CPV antigen for full reaction,then dropped the mixture into the CPV colloidal gold test strip,so according to the highest serum dilution ratios when the test strip line T (line T) vanishes,it was to judge CPV antibodies in serum of the HI titer.This method had been used to detect 86 canine serum samples,at the same time,analyzing and comparing it with traditional hemagglutination inhibition test method.The results showed that the CPV antigen detection test strip was successfully prepared,and the reaction conditions and results of the test strip for detecting the titer of CPV-HI in canine serum were determined.The results indicated that when detecting CPV antigen after the dilution of different ratios of canine serum,the highest serum dilution ratios when the strip line T vanished and the HI titer had positive correlation.The highest dilution ratios of canine serum multiplied by 4 was the HI titer.The results of two methods had 90.7% consistency.This experiment established the colloidal gold immune chromatography test strip for the detection of CPV-HI titers method initially.This CPV-HI detection provided a simple and fast test method for the effect evaluation of CPV vaccine immune.     

犬细小病毒血凝抑制抗体效价胶体金免疫层析检测方法的建立及应用

试验旨在利用胶体金免疫层析技术建立快速检测犬血清中犬细小病毒（canine parvo virus, CPV）血凝抑制（haemagglutination inhibition, HI）抗体效价的方法,用于CPV疫苗免疫效果评价。采用双抗体夹心法,以抗CPV血凝相关抗原的单克隆抗体制备CPV抗原检测试纸条;将犬血清进行不同比例系列稀释后,分别与定量CPV抗原充分反应,滴入CPV胶体金试纸条,根据试纸条检测线（test line,T线）消失时的血清最高稀释倍数判断血清中CPV抗体的HI效价;用此方法检测86份犬血清样品,并与传统血凝抑制试验方法进行分析比较。结果显示,成功制备CPV抗原检测试纸条,确定了试纸条检测犬血清CPV-HI效价的反应条件和结果判定标准。结果表明,在检测不同稀释倍数犬血清反应后的CPV抗原时,能使试纸条T线消失时的血清最高稀释倍数与HI效价具有正相关性,犬血清最高稀释倍数乘以4即为HI效价;两种方法的符合率达90.7%。本试验初步建立了胶体金试纸条检测CPV血凝抑制效价的方法,为检测CPV-HI效价提供了一种操作简单、快速的试验方法,可用于CPV疫苗免疫效果评价。     

犬用四联疫苗免疫试验

用CDV弱毒、CPV弱毒、CAV 2型弱毒和ERA弱毒成功研制了犬用四联疫苗。该四联疫苗安全可靠 ,免疫原性强 ,免疫期 6个月以上。该四联疫苗免疫剂量为 3mL/只 ,CDV、CPV和CAV 2型和ERA弱毒疫苗效价为 10 6 0 ～ 10 7 5,应用接种 30万只犬 ,抗体阳转率为 90 %。     

犬细小病毒病免疫预防研究进展

犬细小病毒病是由犬细小病毒（Canine parvovirus,CPV）引起的一种高度接触性传染病,发病犬临床表现为严重的出血性肠炎和心肌炎综合征。本病全年均有发生,但相对集中在2月～6月份,2月龄～4月龄幼犬易感性最强。病毒主要随污染的饲料、饮水经消化道进入机体引起发病。有些病犬康复后,仍可长期通过粪便排毒,成为本病的重要隐性传染源。一旦发生CPV感染,除早期应用抗血清及对症疗法有效外,无其他特效疗法,中晚期病例多预后不良,因此,必须依靠免疫预防控制该病的发生。本病的预防办法主要是定期进行免疫接种,接种途径主要是肌肉注射。免疫预防要取得满意效果,平时就必须严格执行防疫措施,但免疫的程序与时机等因素都会影响免疫效果。文章就以上内容对犬细小病毒感染免疫预防研究进展做了综述。     

Non-Structural protein 1 (NS1) gene of Canine Parvovirus-2 regresses chemically induced skin tumors in Wistar rats

The Non-Structural protein 1 of Canine Parvovirus-2 (CPV2.NS1) plays a major role in viral cytotoxicity and pathogenicity. CPV2.NS1 has been proven to cause apoptosis in HeLa cells in vitro in our laboratory. Here we report that CPV2.NS1 has no toxic side effects on healthy cells but regresses skin tumors in Wistar rats. Histopathological examination of tumor tissue from CPV2.NS1 treated group revealed infiltration of mononuclear and polymorphonuclear cells with increased extra cellular matrix, indicating signs of regression. Tumor regression was also evidenced by significant decrease in mitotic index, AgNOR count and PCNA index, and increase in TUNEL positive apoptotic cells in CPV2.NS1 treated group. Further, CPV2.NS1 induced anti-tumor immune response through significant increase in CD8⁺ and NK cell population in CPV2.NS1 treated group. These findings suggest that CPV2.NS1 can be a possible therapeutic candidate as an alternative to chemotherapy for the treatment of cancer.     

Systemic and local intestinal antibody response in dogs given both infective and inactivated canine parvovirus

Systemic and local immune responses were evaluated in dogs given infective canine parvovirus (CPV) and 2 administrations of inactivated CPV 6 months later. Before the inactivated CPV was given, a jejunal cannulation was performed on the animals. During infective CPV administration, concentrations of class-specific copro- and sero-immunoglobulin (Ig)G, IgA, and IgM were determined by enzyme-linked immunosorbent assay. High concentrations of both copro- and sero-IgM, as well as a moderate increase in the concentrations of sero-IgG and copro-IgA, were detected within 3 days after experimental challenge. Hemagglutination inhibition titers correlated with both serum anti-CPV IgG and IgM early and serum anti-CPV IgG in the later states of infection. After 2 oral administrations of inactivated CPV, class-specific jejuno-, copro-, and sero-IgG, IgM, and IgA anti-CPV antibodies also were determined by enzyme-linked immunosorbent assay. High concentrations of jejuno-IgM and moderate levels of jejuno-IgG and IgA were found. Copro-IgM was not detected in the feces; however copro-IgG and IgA were. Also, no correlations were found to exist when sero-IgM and IgG were compared with jejuno- and coproantibody concentrations throughout the experimental period. Thus, it appears that the immune responses to CPV include both a secretory component of the intestinal mucosa and a systemic component of peripheral lymphoid tissues. Application of the modified Witzel's enterostomy to this study of local intestinal immunity proved beneficial. The technique was well tolerated by experimental animals and allowed for simple, multiple sample collections of intestinal contents for virus and antibody determinations with apparently minimal alterations in the luminal environment.     

Application of a dot enzyme-linked immunosorbent assay for evaluation of the immune status to canine parvovirus and distemper virus in adult dogs before revaccination.

A growing body of literature has been published indicating that the current practice of annual vaccination of dogs may not be beneficial and in some cases may even be harmful. A number of publications have proposed assessing the immune status of dogs before annual revaccination. In this study the usefulness of a commercially available dot-ELISA kit was evaluated to determine the duration IgG antibody titers to canine parvovirus (CPV) and canine distemper virus (CDV) in 158 dogs vaccinated at least one year ago. Overall, the percentage of dogs with protective antibody titers to both CPV and CDV was 84%. The percentage of dogs with borderline antibody titers was 11% for CPV and 10% for CDV. Four percent of the dogs had no detectable antibody to CPV and 6% had no antibody to CDV. The results reported here are in good agreement with other studies measuring IgG antibody levels. It is concluded that the kit offers veterinarians the opportunity of determining antibody titers and revaccinating only those pets whose antibody titers to specific diseases have waned.     

Prevalence of positive antibody test results for canine parvovirus (CPV) and canine distemper virus (CDV) and response to modified live vaccination against CPV and CDV in dogs entering animal shelters

Canine parvovirus (CPV) and canine distemper virus (CDV) infections are relatively common in animal shelters and are important population management issues since the immune status of incoming dogs is usually unknown. This study aimed to determine the prevalence of positive antibody test results for CPV and CDV in incoming dogs aged ≥ 4 months and to measure antibody response over 2 weeks following vaccination with a modified live vaccine (MLV). Dogs aged 4-24 months entering an adoption-guarantee shelter (Shelter 1, n=51) and aged ≥ 4 months entering a limited admission shelter (Shelter 2; n=51) were enrolled. Dogs from Shelter 1 had been vaccinated with MLV at a municipal shelter 5 days before enrollment, whereas dogs from Shelter 2 had no known history of vaccination at enrollment. Sera were obtained on day 1, immediately prior to CPV/CDV MLV, and tested using an in-clinic ELISA kit to detect CPV/CDV antibodies. Dogs negative for CPV and/or CDV were retested at day 6-8 and those dogs still negative at day 6-8 were retested at day 13-15. Prior to CPV/CDV MLV on day 1, more dogs tested positive for CPV (Shelter 1 - 68.6%; Shelter 2 - 84.3%) than for CDV (Shelter 1 - 37.3%; Shelter 2 - 41.2%). On day 1, prior to MLV, all spayed/neutered animals tested CPV antibody-positive (n=17/102) and CPV antibody-positive dogs were older than serologically negative dogs (Shelter 1, P=0.0029; Shelter 2, P=0.0042). By day 13-15, almost all dogs were CPV antibody-positive (Shelter 1 - 97.9%; Shelter 2 - 100.0%) and CDV antibody-positive (Shelter 1 - 93.8%; Shelter 2 - 97.8%). MLV induces protective antibody titers against CPV/CDV in almost all dogs after 13-15 days.     

A modified live canine parvovirus vaccine. II. Immune response

The safety and efficacy of an attenuated canine parvovirus (A-CPV) vaccine was evaluated in both experimental and in field dogs. After parenteral vaccination, seronegative dogs developed hemagglutination-inhibition (HI) antibody titers as early as postvaccination (PV) day 2. Maximal titers occurred within 1 week. Immunity was associated with the persistence of HI antibody titers (titers greater than 80) that endured at least 2 years. Immune dogs challenged with virulent CPV did not shed virus in their feces. The A-CPV vaccine did not cause illness alone or in combination with living canine distemper (CD) and canine adenovirus type-2 (CAV-2) vaccines, nor did it interfere with the immune response to the other viruses. A high rate (greater than 98%) of immunity was engendered in seronegative pups. In contrast, maternal antibody interfered with the active immune response to the A-CPV. More than 95% of the dogs with HI titers less than 10 responded to the vaccine, but only 50% responded when titers were approximately 20. No animal with a titer greater than 80 at the time of vaccination became actively immunized. Susceptibility to virulent CPV during that period when maternal antibody no longer protects against infection, but still prevents active immunization, is the principal cause of vaccinal failure in breeding kennels where CPV is present. Reduction, but not complete elimination, of CPV disease in large breeding kennels occurred within 1-2 months of instituting an A-CPV vaccination program.