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1.
Monoclonal antibodies against hog cholera and bovine viral diarrhoea viruses were assayed on organ tissue sections of experimentally infected animals. The animals had been infected simultaneously with both viruses. The antibodies were tested using an indirect immunofluorescence test and an indirect enzyme immunoassay with a biotin/streptavidin/peroxidase detection system. A polyclonal hyperimmune serum was used as a control in direct immunofluorescence tests. Both techniques based on monoclonal antibodies were more sensitive and more specific than the conventional test, the enzyme immunoassay being more sensitive than the immunofluorescence test. Small amounts of BVD viral antigen were demonstrable with monoclonal antibodies in most organ tissues.  相似文献   

2.
In the absence of detectable cytologic changes in hog cholera virus-infected tissue culture cells, hog cholera viral antigen was readily detected by immunofluorescence. The ability to detect hog cholera viral antigen by this method allowed for determination of infectivity titers and also for titration of homologous antibody. Immunofluorescence made possible the identification, in tissue culture, of hog cholera virus from blood, serum, and spleen extracts of experimentally infected swine. Further applications of this method and its limitations are being investigated.  相似文献   

3.
Natural infection of pigs with bovine viral diarrhea virus (BVDV) through contact with infected cattle has caused problems in diagnosing hog cholera (HC). Low cross-reacting serum antibody titers against HC caused by BVDV infection were found in clinically normal pigs as well as those suspected of having HC. Bovine viral diarrhea virus was isolated from specimen tissues and initially identified as HC virus (HCV), using the fluorescent antibody cell culture technique. Additional cell cultures, as well as pig and calf trials, were necessary to identify it as BVDV. The isolate caused clinical signs of illness in the calves, whereas the pigs remained healthy. Bovine viral diarrhea virus may be detected in tissue sections or isolated in cell cultures and confirmed as HCV, using the HC fluorescent antibody conjugate. Laboratories performing the neutralization test for HC should use discretion when interpreting HC titers unless BVD titers are determined on the same serums.  相似文献   

4.
将 2 0头 9月龄左右猪瘟、伪狂犬、猪繁殖与呼吸障碍综合征抗原、抗体阴性猪分成 6组 ,分别利用猪细小病毒(PPV)、猪伪狂犬病毒 (PRV)和猪繁殖与呼吸障碍综合征病毒 (PRRSV)单独或混合感染。 7d后连同对照猪 4头 ,免疫接种猪瘟兔化弱毒疫苗 (HCL V) ,13d后连同 4头阴性对照猪一起攻击猪瘟石门强毒。整个试验期间分别每天测温 ,观察临床症状 ,每周采集扁桃体和血样做各种病毒抗原及抗体检测。结果表明 ,非猪瘟病毒感染 7d后 ,所有各组猪均从体内检测到了相应感染的病原 ,表明 3种非猪瘟病毒感染成功。在攻击猪瘟石门强毒后 2周 ,感染了非猪瘟病毒后接种 HCL V疫苗的 4个免疫组 12头猪除 1头外 ,11头全为猪瘟病毒 (HCV)抗原检测阳性 ,且多呈强阳性 ;而单一 HCL V疫苗免疫组在猪瘟强毒攻击后检测不到 HCV;所有 HCL V疫苗免疫猪均存活 ,而非免疫对照组 4头猪全部在攻毒 16 d内死亡。  相似文献   

5.
The antibodies in serum samples from an outbreak of low-virulent hog cholera in Spielbach, West Germany, 1966, as well as serum samples from pigs inoculated with hog cholera (HG) virus and bovine viral diarrhea (BVD) virus, respectively, were examined by means of 3 different methods:
  1. A modified direct complement fixation (GF) test,
  2. A peroxidase-linked antibody (PLA) assay based on microplates with fixed, viral-antigen containing cells,
  3. A neutralization assay carried out in microplates using the “chessboard” principle and read by means of the peroxidaselinked antibody (NPLA) assay.
A good correlation was found in their ability to detect the antibodies. Generally neutralizing antibodies could be found 2 weeks after inoculation. By CF and PLA antibodies could be detected at the same time or up to 2 weeks later. All sera were tested by the 3 methods against both HG viral antigen and BVD viral antigen. HC-antibodies could not be distinguished from BVD-antibodies by CF but to a certain degree by PLA. BVD-antibodies could to a certain degree be distinguished from HG-antibodies by CF but not by PLA. This means that CF and PLA together provide a good possibility for differentiation between the two types of antibodies. NPLA could to a high degree of reliability distinguish between HG-antibodies and BVD-antibodies.  相似文献   

6.
A liquid-phase enzyme-linked immunosorbent assay (ELISA) was developed for the detection of vesicular stomatitis virus (VSV) types New Jersey (VSV-NJ) and Indiana subtype Indiana I (VSV-IND1) antibodies in the sera of naturally and experimentally infected cattle, horses and swine. Four different VSV preparations were compared for use as antigen in the ELISA: virus used in neutralization tests, complement-fixation antigen, immunodiffusion ager gel antigen and viral glycoprotein. Comparative antibody titration results of virus neutralization (VN) and ELISA showed no statistically significant difference between serum titers obtained with the four antigens to VSV-NJ (P=0.21) and VSV-IND1 (P=0.14). The viral glycoprotein antigen was incorporated in the ELISA system because it is non-infectious and induces neutralizing antibodies. The reliability and variability of the ELISA was determined by testing 516 bovine, equine and swine sera which originated from areas free of vesicular stomatitis, and by testing 186 sera from areas where outbreaks occur. ELISA and VN results were correlated (P < 0.001 for both serotypes), and no statiscafically difference was found between replications of the tests. The ELISA allows the testing of a larger number of sera in a shorter time than is possible with the VN test and it can be used in diagnostic laboratories in VSV-free areas for the support of epidemiological surveillance programs.  相似文献   

7.
A soluble antigen present in infectious tissue culture fluids was separated from the infective virus particle by ultracentrifugation of two serologically related strains of bovine viral diarrhea viruses, NADL-MD and Oregon C24V.

Neutralizing antibodies against the two viruses were absent in four hog cholera antisera, but present in significant titer in the commercially prepared antiserum. Precipitin tests utilizing the agar double diffusion technique formed a single line of identity between the concentrated soluble antigen of both viruses and NADL-MD and hog cholera antisera. No lines were observed using concentrated virus pellet and noninfected BEK cell antigens or control SPF calf and swine sera.

  相似文献   

8.
In calves inoculated with bovine viral diarrhea (BVD) viruses and soluble antigen, the complement-fixing (CF) antibodies appeared before serum-neutralizing (SN) antibodies and remained at high levels throughout the test period. A rapid rise in SN antibodies occurred after challenge with homologous virus with no apparent effect on CF antibody levels.

The CF antibody responses in calves infected with cytopathogenic NADL-MD and noncytopathogenic CG-1220 viruses were similar whereas SN antibody responses indicated strain specificity by reciprocal cross-neutralization tests.

The CF antibody levels in 5 hog cholera (HC) antisera were assayed using the soluble antigen of NADL-MD BVD virus. No demonstrable SN antibodies were present in four HC antisera tested against NADL-MD virus, but a significant titer was present in the commercially prepared antiserum.

Virus was reisolated from animals infected with BVD viruses by buffy coat culture technique during 3 weeks postinoculation, even when significant levels of CF and SN antibodies were present.

  相似文献   

9.
Clinical signs and lesions can sometimes provide the basis for a presumptive diagnosis of hog cholera (HC). However, an accurate diagnosis requires laboratory testing. The usual procedure for the detection of viral antigen is the examination of cryostat sections stained with fluorescein-conjugated HC antiserum. A more definitive technique is isolation of the virus in PK-15 cell cultures and identification of the viral antigen in cells using an HC fluorescent antibody conjugate. As bovine viral diarrhea (BVD) virus will cross-react with HC virus, isolation must be confirmed by the comparison of BVD and HC staining or, preferably, by the use of monoclonal antibodies that can differentiate between HC and BVD viruses. Hog cholera surveillance must rely on serology. The fluorescent antibody virus neutralization (FAVN) test is the classical technique, and HC and BVD antibody can usually be differentiated if HC-positive serum samples are tested against both viruses. Recently the enzyme-linked immunosorbent assay (ELISA) and peroxidase-labeled antibody tests have become the commonly used techniques.  相似文献   

10.
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11.
A serodiagnostic ELISA utilizing the recombinant nucleoprotein (rN protein) of transmissible gastroenteritis virus (TGEV) was developed, and evaluated by examining a panel of 141 virus neutralization (VN) positive and 101 negative sera. The rN protein-based ELISA (rnELISA) appeared to be highly sensitive and specific (98.6% and 98.0%, respectively) when it was compared to the VN test. The result was similar to that of an ELISA based on purified viral antigens with showing good correlation (R=0.829). No cross-reaction was detected with antisera against porcine epidemic diarrhea virus, hog cholera virus, type A rotavirus, pseudorabies virus and swine vesicular disease virus in this ELISA. The rnELISA can be an alternative for the diagnosis of TGE with a great advantage in antigen preparation.  相似文献   

12.
Monoclonal antibodies to bovine viral diarrhea virus (BVDV) were examined for binding with a large number of North American BVDV isolates and eight strains of the serologically related pestivirus, hog cholera virus (HCV). No single BVDV monoclonal antibody reacted with all BVDV isolates. The most cross-reactive monoclonal antibody was an anti-p80/p125 antibody which showed a positive reaction with 173 of 180 (96%) North American isolates. From a fewer number of isolates tested, one anti-gp53 monoclonal antibody also showed a high cross-reactivity (94%). All BVDV isolates showed a positive reaction with at least one of the seven monoclonal antibodies in the panel. Thus, the results indicated that a pool of these monoclonal antibodies may be used in place of polyclonal antisera for the detection of BVDV contamination of cell lines or for virus isolation. For HCV, all three anti-p80/p125 monoclonal antibodies reacted positively with all eight virus strains. In contrast, none of the anti-gp53 monoclonal antibodies were reactive to HCV strains. Thus, the anti-gp53 monoclonal antibodies may be useful for distinguishing between usually innocuous BVDV infections and the highly significant HCV infections in swine for foreign animal disease surveillance.  相似文献   

13.
Thirty-three pestivirus strains were grown in cell culture and characterized by immunostaining with 19 monoclonal antibodies (MAbs) raised against hog cholera virus (HCV), with 42 MAbs against bovine viral diarrhoea virus (BVDV) and with 13 MAbs against border disease virus (BDV). Seven MAbs reacted with all pestivirus strains tested, eight MAbs detected only the seven HCV strains, three detected only the 16 BVDV strains. No MAb was found that was specific for BDV. BVDV and BDV strains were broadly cross-reactive with the MAbs, indicating a close relationship between these two species, whereas HCV strains were characterized as distinct from BVDV and BDV.  相似文献   

14.
A panel of 30 monoclonal antibodies was defined and characterized with respect to the binding capacity in immunoperoxidase assay to different strains of pestivirus. Using the panel it was possible to identify specifically all strains and isolates of hog cholera virus, hog cholera vaccines derived from 'C' strains, and most strains of bovine viral diarrhoea/border disease (BVD/BD) viruses (including those isolated from pigs). A small proportion of BVD/BD isolates from pigs and ruminants reacted only with the monoclonals specific for pestivirus group antigen. It is recommended that monoclonal typing methods be introduced into official procedures for the diagnosis of hog cholera/classical swine fever.  相似文献   

15.
Specific staining of antigen within bovine embryo kidney tissue culture cells, infected with either Oregon C24V or NADL-MD bovine viral diarrhea virus, was accomplished using fluorescein-conjugated swine anti-hog cholera or bovine antiviral diarrhea globulin. Also specific staining of antigen within pig kidney tissue culture cells, infected with hog cholera virus, was accomplished using the same two types of conjugates. Specificity was confirmed by appropriate controls.  相似文献   

16.
猪瘟病毒石门株基因组cDNA片断的扩增与克隆测序   总被引：4，自引：0，他引：4  
以猪瘟病毒（ＨＣＶ）中国石门株强毒的血毒、细胞毒及Ｃ系兔化弱毒的细胞毒中提取的总ＲＮＡ为模板，应用逆转录─聚合酶链式反应（ＲＴ一ＰＣＲ），在合成的两对ＨＣＶ特异引物引导下，扩增出与预计大小一致的４４１和３００ｂｐ两个核酸片断。寡核苷酸探针杂交证实确为ＨＣＶＰ_８０和ｇｐ４４／４８蛋白编码区特异性的ｃＤＮＡ片断。扩增片断克隆入ｐＵＣ_１９和ｐＢＳＫ（＋）中，并对ＨＣＶ石门株Ｐ_８０区ｃＤＮＡ片断进行测序分析，其与国外Ａｌｆｏｒｔ、Ｂｒｅｓｃｉａ株同源性分别为９０．６％和９４．６％。氨基酸水平上同源性高达９８．４％。为抗猪瘟病毒Ｐ_８０区核酶的设计和应用提供了依据。  相似文献   

17.
猪繁殖与呼吸综合征病毒感染抑制猪瘟疫苗的免疫应答   总被引：23，自引：3，他引：23  
对20日龄SPF猪和20日龄猪繁殖与呼吸综合征（PRRS）血清阳性猪，人工感染猪繁殖与呼吸综合征病毒（PRRSV）北京分离株（BJ－4）后48h接种猪瘟疫苗，利用ELISA方法检测仔猪针对PRRSV和猪瘟疫苗的体液免疫，利用MTS法检测仔猪外周血单核细胞对有丝分裂原ConA的刺激反应。结果表明，不论是SPF仔猪还是带有PRRSV抗体的仔猪，在鼻内接种PRRSV后48h接种猪瘟疫苗，其对猪瘟疫苗的抗体反应显著低于对照组，对有丝分裂原ConA的刺激反应也下降。由此说明，PRRSV感染使仔猪对猪瘟疫苗的免疫应答受到抑制。  相似文献   

18.
A collection of 90 field isolates of hog cholera virus (HCV) was used to test the specificity of four hybridoma cell lines secreting monoclonal antibodies against pestiviruses. Reaction of virus isolates and monoclonal antibodies was controlled by an indirect immunofluorescence assay (IFA). Two monoclonal antibodies which had been generated against HC virus strain "Alfort 187" were reactive only with HCV field isolates and an HCV reference strain but not with bovine viral diarrhoea virus (BVDV) reference strains. Two other monoclonal antibodies (generated against BVDV, strain NADL) reacted only with BVDV reference strains but not with HCV field isolates, although with 3 of these strains focal reactions involving only a few cells were detected. The ability to discriminate between both viruses is a diagnostic need which may be fulfilled by these monoclonal antibodies.  相似文献   

19.
Thirty-one bovine viral diarrhoea virus (BVDV) or border disease virus (BDV) strains and 94 hog cholera virus (HCV) strains were grown in cell culture, and characterized by immunostaining with 13 monoclonal antibodies (MAbs) and one polyclonal serum (PAb) against HCV. All 125 strains were recognized by the PAb. None of the BVDV or BDV strains were detected by the 13 MAbs. Seven MAbs detected all 94 HCV strains. Six other MAbs detected heterogeneity among and within HCV strains. The MAbs are useful tools in differentiating between HCV and BVDV infections in pigs, and can also be used to differentiate infections induced by HCV field strains from infections induced by the "Chinese" strain of vaccine virus.  相似文献   

20.
应用FA及PPA-ELISA技术对猪瘟的检测   总被引：3，自引：0，他引：3  
用免疫荧光抗体诊断技术及单克隆抗体纯化酶联免疫吸附试验诊断技术对疑为猪瘟病毒感染猪进行了检测 ,重点阐述了 2种方法的技术原理和操作过程。通过用免疫荧光抗体诊断技术检测了 5份病料 ,其中有 2份为阳性 ,阳性率为 40 % ;用单克隆抗体纯化酶联免疫吸附试验诊断技术检测了猪瘟血清抗体 ,在 40头母猪血清样品中 ,猪瘟弱毒抗体效价 OD值较高 ,有 1 0 0 %的保护率 ,但是发现母猪群中有 1 0 %隐性猪瘟感染 ,其强毒抗体效价 OD值大于 0 .5,体内带有猪瘟病毒。结果及过程表明此两种诊断方法检测快速、鉴别准确、分辨率高  相似文献   

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