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1.
An indirect immunofluorescence (IF) test was developed to detect bovine leukemia virus (BLV) antigen expression in infected sheep lymphocytes, using monoclonal antibodies anti BLV-major envelope glycoprotein gp51. Peripheral blood lymphocytes were cultivated for 48 h in presence of phytohemagglutinin (PHA) (50 μg/ml), and then fixed with acetone. The cells were assayed for the IF test. All experimentally infected sheep were positive with this test.  相似文献   

2.
Antibody titers to protein (p) and glycoprotein (gp) antigens of bovine leukemia virus were studied by the immunodiffusion test in two groups of cattle. One group showed evidence of enzootic bovine leukosis (BL+ cattle), whilst the second group, possessing antibodies to gp antigen, were adjudged as subclinical cases (BL?). All 165 BL? and 97% (108/111) BL+ cattle possessed antibodies to gp antigen but those to p antigen were evident in only 67% BL? cattle in contrast to 89% in BL+ cattle. Mean antibody titers to both antigens were statistically higher in BL+ cattle.  相似文献   

3.
Twenty-seven cows with antibodies to bovine leukemia virus were bled before, during and after calving. All serum samples were tested quantitatively for bovine leukemia virus antibodies using both the agar-gel immunodiffusion test with a glycoprotein antigen and the radioimmunoprecipitation assay with an internal p24 protein antigen. A significant fall (P less than 0.001) in bovine leukemia virus-antibody titer was demonstrated with both tests at the time of calving, with a subsequent rise in antibody titer within one month of parturition. Bovine leukemia virus antibodies were not detectable using the agar-gel immunodiffusion test in two of these cows at the time of calving.  相似文献   

4.
A sensitive and specific radioimmunoassay for the major internal antigen of bovine leukemia virus has been applied to detecting this protein in cultured lymphocytes of infected cattle. The specificity inherent in this assay offers obvious advantages over a previously described syncytium induction assay for infectious bovine leukemia virus, because false positive reactions due to other viruses such as bovine syncytial virus are avoided. Investigations of various culture conditions indicated that maximal amounts of antigen had been produced after incubation for 72 hr at 37°C. Lymphocyte concentrations of 106?5×107 cells/ml gave satisfactory results. Tests of cultured lymphocytes from bovine leukemia virus infected or bovine leukemia virus-free cattle indicated a comparable sensitivity between the radioimmunoassay and syncytium induction assay in the detection of bovine leukemia virus infections.  相似文献   

5.
Six cattle persistently infected with bovine virus diarrhoea virus (BVDV) and seronegative, and two control, virus negative seropositive cattle were inoculated with lymphocytes infected with bovine leukosis virus (BLV). The two controls produced a normal immune response to BLV, developing antibodies at four and five weeks after inoculation. Two of the six cattle persistently infected with BVDV developed a strong antibody response by six weeks after inoculation with BLV. Four developed a depressed response to BLV, characterised in three by a 'hooking' reaction in the immunodiffusion test which persisted in successive bleedings but was interspersed occasionally by a weak positive reaction. In one of these animals, a series of 'hooking' reactions was followed by a number of negative results. The fourth animal remained serologically negative until 16 weeks after inoculation when a 'hooking' reaction was observed followed by a series of negative results. BLV was isolated from all the cattle persistently infected with BVDV at 42 or 58 weeks after inoculation regardless of whether the serum samples gave negative, 'hooking', weak positive or positive reactions in the immunodiffusion test. BLV was consistently isolated from the nasal secretions of a steer which was BVDV negative but seropositive. The possibility of decreased immune responsiveness to BLV in animals persistently infected with BVDV should be considered when formulating regulations governing the testing of animals for freedom from BLV.  相似文献   

6.
Four tests for detection of antibodies to bovine leukemia virus (BLV) were compared. The sera that were tested came from cattle in naturally infected commercial dairy herds, cattle that were infected under experimental conditions, and cattle in an isolated BLV-free herd. The tests that were compared included a radioimmunoprecipitation assay (RIA) with p24 antigen, a RIA with glycoprotein (gp) antigen, an agar-gel immunodiffusion (AGID) test with gp antigen, and a virus-neutralization (VN) test that was based on inhibition of BLV-induced syncytia in cell culture. Results of the 4 serologic tests agreed for 96.8% of the sera from cattle in commercial herds. The gp RIA detected the greatest number of positive sera (188); it was followed in turn by the p24 RIA (187), the VN test (183), and the AGID test (176). The gpd RIA titers of the 12 sera that gave negative AGID results were 175 or less. In RIA, the percentage of precipitation of labeled antigen by positive sera was almost always higher with gp antigen than with p24 antigen. Satisfactory sensitivity in the p24 RIA required the acceptance of a low level of antigen precipitation, 15%, as a positive test. In the gp RIA, however, almost all positive sera precipitated at least 50% of the labeled antigen. Nonspecific precipitation of antigen in the RIA by sera from BLV-free cattle ranged from 4% to 10%. Examination of sequential serum samples from 17 experimentally infected cattle showed that BLV antibody was first detected 2 to 8 weeks after inoculation. In 9 cattle, seroconversion was detected simultaneously by all of the tests. Results from the other 8 cattle indicated that seroconversion could be detected first by p24 RIA, followed by the gp RIA and the VN test. The longest interval between RIA seroconversion and AGID seroconversion was 10 days. Monthly tests of sera from 10 laboratory cattle that were infected by contact exposure showed that 7 animals seroconverted in all tests at the same time. Two cattle were positive first in RIA, but the next month they were also positive in the VN and AGID tests. One animal was positive in the RIA and the VN test for 2 months before antibody was detected by AGID.  相似文献   

7.
A seroepidemiological study of bovine immunodeficiency virus (BIV) and bovine leukemia virus (BLV) infections was conducted in four different cattle herds in Turkey. A total of 300 blood samples were analyzed and 12.3% were found to be positive for anti-BIV p26 antibodies by Western blot analysis and 1.6% positive for anti-BLV gp51 antibodies by an immunodiffusion test. BIV infection was confirmed with the detection of BIV-provirus DNA using the nested polymerase chain reaction. This is the first evidence for the presence of BIV in cattle in Turkey.  相似文献   

8.
Expression of bovine leukemia virus (BLV) antigens in vivo has not been shown. After BLV infection, however, production of antibodies directed towards BLV proteins (e.g. gp51) can be easily demonstrated. Thus, production of BLV proteins has to take place somewhere in infected cattle. Tissues and organs of experimentally infected cattle were fixed in acetone and embedded in paraffin. Monoclonal antibodies directed to gp51 were used to demonstrate BLV expression immunohistologically by the peroxidase-antiperoxidase (PAP) method. The same samples were also used to demonstrate a tumor associated antigen (TAA) employing a monoclonal antibody. Our results indicate that very few cells, found in the intestinal mucosa, produce gp51 in vivo. The expression of TAA, however, increases significantly shortly after infection with BLV and remains high throughout life.  相似文献   

9.
The purpose of this study was to examine rates of natural in utero infection with bovine leukemia virus for association with breed, sex, dam age, dam parity and time of maternal seroconversion. Analyses conducted for breed and sex, dam age and parity and time of maternal seroconversion were the FUNCAT procedure for categorical data, Wilcoxon Rank Sums test and Fisher's exact test, respectively. A total of 223 calves born between July 1979, and September 1980, to cows infected with bovine leukemia virus in the University of Florida Dairy Research Unit herd were tested for detectable bovine leukemia virus antibodies prior to the consumption of colostrum. Sera were tested for antibodies by agar-gel immunodiffusion and radioimmunoprecipitation using the glycoprotein-51 antigen. In a group of 125 calves in which in utero infection could be confirmed through serological follow-up (group A), eight calves (6.4%) had precolostral bovine leukemia virus antibodies. For all 223 calves (group B), 18 (8.1%) had detectable bovine leukemia virus antibodies. For calves in group A, no associations were detected between precolostral bovine leukemia virus antibodies and breed (p = 0.66), dam age (p = 0.86), dam parity (p = 0.83), or time of maternal seroconversion to bovine leukemia virus (p = 0.50). However, precolostral bovine leukemia virus antibodies were found in 17.4% of the males and 3.6% of the females in group A (p = 0.11) and in 12.4% of the males and 3.6% of the females in group B (p = 0.04).  相似文献   

10.
The 135,000 mw glycoprotein (gp135) and the 28,000 mw internal protein (p28) of caprine arthritis encephalitis virus are major viral constituents in precipitin lines formed between crude antigen preparations and sera from infected goats. In testing 307 goat and sheep sera, 118 samples were positive in a gp135 assay and only 82 were positive in a p28 assay. However, some goat sera were found which reacted only with the p28 and therefore testing for antibody against both proteins may be necessary to identify a maximum number of virus infected goats by immunodiffusion.  相似文献   

11.
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12.
This is the first report of serological evidence for bovine immunodeficiency virus (BIV) infection in Argentina. The analysis was performed in 589 dairy bovine sera samples, applying indirect enzyme-linked immunosorbent assay (I-ELISA) using a synthetic antigen (transmembrane peptide, TM) and Immunofluorescent assay (IFA). In this study, 9 dairy herds from 4 Argentinian provinces were evaluated and 12% of the animals tested positive for BIV. Seven of the 9 herds tested were BIV seropositive and the percentage of BIV seropositive animals in the herds ranged from 2% to 42%. Direct detection of BIV provirus applying nested PCR was not conclusive. Antibody detection against bovine leukemia virus (BLV) in all sera was also performed applying immunodiffusion (ID) assay and 59% resulted seropositive. Statistical analysis of the results was carried out and possible evidence of association between BIV and BLV infection was considered. Future studies should be performed including local field isolates strains of BIV.  相似文献   

13.
An estimated weighted-regression method was used to describe the decay of colostral bovine leukemia virus (BLV) antibodies in the calf, as measured by agar-gel immunodiffusion with glycoprotein antigen. The prediction equation, based on 473 observations from 130 animals, was log10 inverse titer = 1.29 -0.012 age (days). The half-life of BLV antibodies was estimated to be 25.8 days. Ages at which colostral antibodies were last detected were between 51 and 187 days. Normal limits of antibody decay were estimated and used to identify virus-induced active antibodies in calves during the colostral antibody period. Calves known to be infected were identified between 2 and 180 days of age, using 95% limits. Application of this procedure for the early serologic detection of BLV-infected calves in eradication or control programs is discussed.  相似文献   

14.
A trial was performed with heifers at the age of six to seven months. The animals were experimentally infected with the lymphocytes of a virus-productive donor. Infection was produced in all the nine cases, as demonstrated by means of the positive syncytial test. As indicated by the results of the trial, the antibodies to the enzootic bovine leucosis virus (BLV) were produced soon after experimental infection. A high sensitivity of the serum-neutralization test and the ELISA method was demonstrated in this connection: by these methods, the antibodies were identified already two to three weeks after experimental infection whereas by the immunodiffusion test they could be detected only after five weeks. Twenty-four animals were exposed to natural contact infection. Within 270 days of the trial, the disease after contact was recorded only in one heifer out of the four that were in close contact with the experimentally infected animals. In this case, as compared with experimental infection, the antibodies were produced much later--after 85 to 93 days. Leucosis was recorded in none of the remaining animals. The reasons why such a favourable result was obtained were the thorough disinfection of the stables after blood collections and the strict observance of the aseptic conditions. The results of experimental infection in three cows were identical with those obtained in young cattle. In the experimentally infected dairy cows, antibodies in milk were determined by the ELISA method. As found, in milk the antibodies to BLV appear two to three weeks later than they do in serum. The ELISA method of BLV antibody detection can be used for the identification of infected animals in herds where enzootic bovine leucosis occurs.  相似文献   

15.
Cocultivation of equal numbers of cells from a fetal lamb kidney line infected with bovine leukemia virus and African green monkey (Vero) cells results in the rapid production of syncytia. The effect was blocked or inhibited by serum containing antibodies to bovine leukemia virus. A serological test based on syncytial inhibition was compared to the agar gel immunodiffusion test and the modified direct complement fixation test for the detection of bovine leukemia virus antibodies in sera from leukosis-free cattle, cases of adult enzootic bovine lymphosarcoma and cattle from herds in contact with enzootic lymphosarcoma. The results showed the syncytial inhibition test to react positively with sera from all cases of adult enzootic lymphosarcoma, but to be much less sensitive than the other tests in detecting bovine leukemia virus antibodies in sera of exposed animals.  相似文献   

16.
A highly specific monoclonal antibody (mAb) directed against envelope protein gp51 and effectively bonding the antigen (Ag) on account of its high affinity from an unpurified Ag preparation was chosen for use in a double-sandwich enzyme immuno-assay (EIA) for diagnosis of bovine leukaemia virus (BLV). The epitopes recognised in bovine sera by the gp51-specific antibodies were at the same time properly exposed. Some parameters of major importance to testing were optimised (Ab and Ag quantities, dilution of bovine sera for testing). Preliminary testing of the double-sandwich EIA on selected bovine sera and comparison with both the immunodiffusion test and anti-BLV EIA confirmed its good diagnostic specificity and sensitivity. Hence, this double-sandwich EIA, developed by means of an mAB against gp51, on account of the possibility to use as Ag culture supernatant of the FLC cell line, is a sensitive, low-cost alternative to the anti-BLV EIA Dessau MTP which had so far been used. The double-sandwich EIA is recommended for use in final sanitation for its high analytical and diagnostic sensitivity.  相似文献   

17.
Short-term lymphocyte cultures from bovine leukemia virus (BLV)-infected cattle were tested for BLV-associated antigens at various times after incubation. Several immunologic methods were used, including fluorescent antibody tests, immunodiffusion, and radial immunodiffusion. Antigens were not detected in uncultured lymphocytes. The BLV-associated antigens were detected as early as 3 hours, with maximum antigen production occurring at 18 to 24 hours after incubation. These results indicate that culturing of lymphocytes in vitro is necessary for the expression of the virus.  相似文献   

18.
A double polymerase chain reaction (PCR) assay has been devised for the direct detection of bovine leukemia virus (BLV). The assay was directly performed on blood leukocytes, avoiding the DNA-purification procedures. The PCR products were identified by gel-electrophoresis and the specificity of the test was confirmed by hybridization with a biotinylated oligonucleotide probe. When testing the sensitivity of PCR, less than eight genome copies of the provirus were detected in the background of two million negative lymphocytes. In a BLV infected herd 22 animals of various age groups were examined by the indirect (serological) diagnostic tests of agar-gel immunodiffusion and indirect ELISA as well as by the direct detection method of PCR. The tests were repeated at monthly intervals on five occasions. When examining the specimens from cows and heifers, a close agreement was found between the results of the various methods. The newborn calves, which were the offspring of BLV infected mothers, were consequently negative in PCR throughout the experimental period. However, in the indirect tests the calves were positive during the first samplings and became negative only around four months of age. Since the indirect tests can not discriminate infection from colostral immunity, PCR proved to be a useful complementary assay for the safe diagnosis of BLV infection in young calves.  相似文献   

19.
In a double-blind study, the commercial agar-gel immunodiffusion test (AGID) was compared with a radioimmunoprecipitation assay (RIA) performed with glycoprotein (gp) antigen for detection of antibodies to bovine leukemia virus. Of 240 sera tested, 115 were from adult cows and 125 were from precolostral calves. Most adult animals were tested within 1 week of parturition. Sera from 74 cattle were positive and sera from 166 cattle were negative by gp RIA. Sensitivity of the AGID, compared with the gp RIA, was 85.1% when the test was read at 48 hours and was 94.6% when read at 72 hours. Specificity increased from 92.2% at 48 hours to 96.4% at 72 hours. Reading the AGID again at 72 hours also clarified most reactions that were questionable at 48 hours due to a haze around the test serum well. Of 3 RIA-positive precolostral calf sera, 2 were AGID-negative and 1 had a questionable reaction by the AGID at 48 hours. Of 5 RIA-positive sera that were AGID-negative at 48 hours, 2 were precolostral calves and 3 were cows tested at parturition. Of 166 RIA-negative reactions, none was falsely positive by the AGID at 48 or at 72 hours.  相似文献   

20.
Bovine leukemia virus (BLV) has a long latency period during which animals are inapparently infected, may spread the disease, and are only detected by serological techniques or by the most cumbersome molecular biology techniques. We have compared techniques for detecting either total antibodies (ELISA), anti-p24 and Gag-related proteins (Western blot), or anti-gp51 (agar gel immunodiffusion, AGID, and syncytia inhibition, SI) in rabbits inoculated experimentally with inocula of variable immunogenicity. The two tests to detect antibodies to gp51 correlated well in sera clearly positive or clearly negative by either one, but correlation was poor in the intermediate groups. All sera positive by AGID were also positive by ELISA, but results did not agree in sera negative by AGID, ELISA proving to be more sensitive. Western blot was a good technique for detecting antibodies against Gag-related proteins. However, no band was identified to clearly correspond to anti-Env-related proteins. As for other retroviruses, testing of animals for infection with BLV should include the detection of antibodies anti-Gag and anti-Env proteins.  相似文献   

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