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1.
The detection of virus-specific immunoglobulin M (IgM) antibodies in acute-phase serum samples offers the possibility of making an accurate and rapid serologic diagnosis. We have developed a solid-phase capture assay that uses murine monoclonal antibodies specific for bovine IgM to separate the whole IgM fraction of a bovine serum sample. The IgM specific for bovine herpesvirus-1 (BHV-1) is then detected by the addition of viral antigen, which in turn is detected by BHV-1-specific monoclonal antibodies conjugated to horseradish peroxidase. A BHV-1 IgM antibody response was detected during the early postinfection period (7-40 days PI). Bovine herpesvirus-1 IgM antibody was not detected in sera taken from 3 animals following dexamethasone-induced viral reactivation. This method compares favorably with viral isolation, antigen detection in the clinical samples, and paired serology in the diagnosis of BHV-1 infection at a herd level.  相似文献   

2.
为了调查新疆地区某规模化奶牛场牛传染性鼻气管炎（IBR）发病情况,通过采集不同生长阶段牛群血清共计362 份,使用牛传染性鼻气管炎病毒gB（IBR-gB）抗体检测试剂盒检测牛传染性鼻气管炎病毒（IBRV）抗体效价,评估该奶牛场IBRV疫苗免疫效果。结果显示,后备牛中犊牛和青年牛IBRV抗体阳性率分别为88.57%（31/35）、75.00%（21/28）;成年母牛中泌乳期母牛、干奶期母牛IBRV抗体阳性率分别为81.46%（145/178）、95.04%（115/121）。阴性数共44 份,可疑数8 份,IBRV抗体平均阳性率为88.38%;结果表明,疫苗接种后,不同生产阶段牛群均可产生不错的抗体保护效果,为奶牛场防控IBR提供依据。  相似文献   

3.
Immunoglobulin concentrations (IgG, IgM, and IgA) in bovine serum, follicular fluid, and uterine and vaginal secretions were determined. The specificities of IgG, IgM, and IgA for virus-neutralizing antibody against bovine viral diarrhea (BVD) and infectious bovine rhinotracheitis (IBR) viruses were also examined. High concentrations of IgG were present in both serum and follicular fluid. The IgG, IgM, and IgA concentrations were low in uterine and vaginal secretions. There was more IgG in the uterus during estrus than at any other time. Virus-neutralizing antibodies against BVD and IBR in serum of cows were mainly the IgG class. There was positive correlation between serum and follicular fluid virus-neutralizing antibody titers fro BVD and IBR. These antibodies may provide some protection for recently ovulated ova.  相似文献   

4.
An enzyme linked immunosorbent assay (ELISA) was applied to the detection of serum antibodies against infectious bovine rhinotracheitis (IBR), parainfluenza-3 (PI3), adenovirus type 3 (adeno 3) and bovine respiratory syncytial (BRS) viruses. Paired serum samples from calves vaccinated with live attenuated virus vaccines were tested. The ELISA compared favorably with the virus neutralization test for detecting serologic responses to IBR, BRS, and adeno 3 viruses or with the hemagglutination inhibition test for PI3 virus. The simplicity, sensitivity and rapidity of the ELISA test makes it a useful tool for immunological studies with respiratory viruses.  相似文献   

5.
An enzyme linked immunosorbent assay (ELISA) using penicillinase was developed in the form of diagnostic kits (Toxokit-G and Toxokit-M) for the detection of IgG and IgM antibodies to Toxoplasma gondii. The performance of both the kits was compared with commercially available diagnostic kits, i.e. Enzygnost-Toxoplasmosis/IgG (Behring Co., Germany), TOXOTEK-G (Flow Lab., U.K.) and Toxoplasma IgM Microassay (Diamedix Corp., U.S.A.) by testing toxoplasma-suspected human serum samples. The results indicate a good reliability between these diagnostic kits. Toxokit-G has 86.66 and 96.05% sensitivity and specificity respectively. The main advantage of Toxokit-G is that the end result can be assessed visually without using sophisticated instruments. Toxokit-M has 100% sensitivity and specificity and test results were not affected by the presence of antitoxoplasma IgG antibodies, rheumatoid factor or antinuclear antibodies.  相似文献   

6.
The immune response to modified live-virus bovine viral diarrhea (BVD) vaccine and infectious bovine rhinotracheitis (IBR) vaccine was examined in calves that had received passive maternal antibodies to these viruses. Blood serum samples from vaccinated and control (nonvaccinated) calves were examined for more than 1 year to determine the rate of decline of passive anti-BVD and anti-IBR antibodies and the effect that vaccination had on these antibody titers. The control calves lost their antibodies to BVD and IBR viruses at the rate of one half their remaining antibody titer every 21 days. Calves serologically responded to BVD vaccine at a time when maternal antibody titers remained between 1:96 and 1:20. However, animals did not seroconvert to the IBR vaccine until maternal antibodies had decreased and become undetectable. Evidence is presented to show that although passive immunity will inhibit IBR vaccination, priming for a secondary response will occur so that on subsequent vaccination, at a time when maternal antibodies have disappeared, the animals will respond anamnestically to IBR vaccination.  相似文献   

7.
West Nile virus (WNV) is a zoonotic pathogen of global importance. In horses with neurological signs, detection of WNV-specific immunoglobulin M (IgM) in serum is widely used to identify clinical cases of WNV encephalitis. Here, we describe the development of two monoclonal antibodies (mAbs) to equine IgM which were used in a WNV IgM-specific enzyme-linked immunosorbent assay (ELISA). Their performance was compared to an established assay based on polyclonal anti-IgM. Check test serum samples from the National Veterinary Service Laboratory (NVSL) were used to evaluate the performance of the three anti-IgM antibodies. The anti-IgM 1-22 mAb correctly identified all NVSL samples. Both the polyclonal antibody and monoclonal anti-IgM 2B-63 identified eight out of ten samples correctly. The three assays were then compared using serum samples from clinically healthy animals (n=33) and horses with neurological signs (n=21). High Spearman rank correlations (0.76-0.86) were found among the ELISA results. Inter-test agreements (weighted kappa) for assay interpretation resulted in strong agreement (0.95) of the results obtained by the mAbs and moderate agreements when monoclonal and polyclonal anti-IgM-based assays were compared. To determine the analytical sensitivities of anti-WNV IgM detection, serial dilutions of WNV IgM-positive serum samples were analyzed. The highest sensitivity was obtained by using the anti-IgM 1-22 mAb to capture IgM from equine serum. In conclusion, the use of monoclonal anti-IgM antibodies can improve the sensitivity of IgM detection in the acute phase of WN disease.  相似文献   

8.
猪圆环病毒2型IgM抗体间接ELISA的建立   总被引：1，自引：0，他引：1  
猪圆环病毒2型(PCV2)是断奶仔猪多系统衰竭综合症的重要原发性病原,其感染的早期检测有助于及时采取防控措施。以重组衣壳蛋白(Cap蛋白)作为包被抗原,通过对反应条件的优化,建立了PCV2 IgM抗体间接ELISA,其最适抗原包被浓度为1.25 μg／mL,最适血清稀释度为1∶100,临界值为0.35。该检测方法与其他5种常见猪疫病阳性血清无交叉反应,特异性良好。批内、批间重复试验的变异系数均在2%~6%,重复性好。对100份临床样本的检测结果表明,该检测方法与国外同类试剂盒相比,敏感性为90.3%,特异性为92.8%,总符合率为92%。试验结果表明,所建立的PCV2 IgM间接ELISA特异性好和敏感性高,适用于PCV2感染早期的流行病学调查。  相似文献   

9.
A total of 1745 healthy cattle from 295 farms in Saskatchewan and Alberta was tested by ELISA for antibodies to four viruses. Antibodies to infectious bovine rhinotracheitis (IBR) virus were found in 37.8% of sera (59.5% of properties), to parainfluenza 3 (PI3) virus in 93.9% of sera (99.7% of properties), to bovine respiratory syncytial (BRS) virus in 78.5% of sera (86.6% of properties), and to bovine viral diarrhea (BVD) virus in 40.6% of sera (66.7% of properties)

The prevalence of PI3 viral antibodies among Saskatchewan cattle was not affected by district of origin, breed, sex, age, or vaccination practices, though BRS viral antibodies appeared less frequent in young, male, and unvaccinated animals. Antibodies to IBR and BVD viruses were less prevalent in the Prince Albert/Tisdale districts and in young, male, and unvaccinated animals, but were more common in Holstein cattle. Antibodies to IBR virus appeared less frequent in Herefords. Antibodies were more prevalent in cattle which had been vaccinated against IBR, BRS, and BVD virus infections.

The relatively small number of cattle sampled from Alberta had a similar prevalence of antibodies to PI3 and BRS viruses to that seen in cattle in Saskatchewan, though IBR and BVD prevalence rates were lower.

  相似文献   

10.
The objective of this study was to evaluate the passive transfer of maternal antibodies to calves following vaccination of pregnant cows with an inactivated Mannheimia haemolytica (MH) and Bovine herpes virus type 1 (IBR) vaccine (Bovilis® MH + IBR). Sixty-two pregnant cows were allocated at random to two groups; one group was retained as a negative control group (T01), while the other group (T02) was vaccinated with Bovilis® MH + IBR on two occasions during their third trimester of pregnancy. Following calving, blood samples were collected from calves for the measurement of serum antibody titres to IBR and MH, with samples collected prior to suckling (Day 0) and on days 5 (±2), 14 (±3), 28, 56, 84, 112, 140, 168, 196, 224, 252 and 280. The group mean IBR blocking percentage remained low for T01 calves (calves born to T01 cows) between days 0 and 224 (range 4.5%–15.4%), while the group mean IBR blocking percentage increased for T02 calves (calves born to T02 cows) from 14.3% on Day 0 to 94.9% on Day 5 and remained significantly higher than T01 calves up until Day 252. The group mean MH titre (Log2) for T01 calves increased after suckling to 8.9 on Day 5, before declining and remaining stable (range 5.0–6.5). The group mean MH titre for T02 calves increased after suckling to 13.6 on Day 5 and then gradually declined; however, it remained significantly higher than T01 calves between days 5 and 140. Outcomes from this study have confirmed that colostral transfer of IBR and MH antibodies to newborn calves was successful and a high level of passive immunity was acquired by calves.  相似文献   

11.
OBJECTIVE: To determine the presence of serum antiretinal antibodies in sudden acquired retinal degeneration syndrome (SARDS) affected dogs and the size of the antigen to which these antibodies bind via the use of enzyme-linked immunosorbent assay (ELISA) and Western blot immunoassays. ANIMALS STUDIED: Serum was collected from 13 dogs affected by SARDS and five dogs with normal ocular examinations. PROCEDURES: All serum samples were subjected to ELISA with saline-soluble canine retinal tissue and Western blot analyses with SDS solubilized normal canine retinal tissue as the antigen. Antirecoverin (23 kDa) and antiheat shock cognate (65 kDa) antibodies were used as positive controls for both procedures. Affinity-purified goat antidog IgG and IgM labeled with horseradish peroxidase were used for all clinical samples and goat antirabbit IgG was used as the secondary antibody for the positive controls. RESULTS: ELISA demonstrated antibody reaction with all samples. Western blot immunoassays identified multiple bands in all canine serum samples, as well as in negative controls. Approximate sizes of the bands were 25 and 50 kDa, corresponding to IgG light and heavy chains, respectively. CONCLUSION: No antiretinal autoantibodies were identified in the serum of dogs affected by SARDS as compared to normal canine patients.  相似文献   

12.
Nine monoclonal antibodies which reacted with equine immunoglobulin (Ig)M and not other equine Ig and serum proteins were prepared. Cells producing antibodies (C 1.9) which precipitated with IgM and bound to staphylococcal protein A were triple-cloned (C 1.9/3.2) and the antibodies further characterized. Monoclonal antibody C 1.9/3.2 reacted with an IgM determinant present on serum IgM from horses of several breeds. Studies with 125I-labeled IgM revealed the presence of this determinant on all IgM molecules. The monoclonal antibody enabled quantitation of IgM in presuckling foal and adult horse sera, using rocket electrophoresis. This procedure was used because presumably it gives a positive precipitation reaction over a wide range of antigen-antibody ratios. The C 1.9/3.2 monoclonal antibody recognized an exposed mu-chain determinant on live B lymphocytes, as determined by immunofluorescence. Also, IgM-containing cells could be identified in acetone-fixed frozen sections of lymphoid tissue. Sera from several other species carry the determinant identified by C 1.9/3.2, suggesting that the reagent may be useful for IgM studies in other species.  相似文献   

13.
钟承  潘玲  刘亚 《中国奶牛》2012,(21):36-38
本试验对蚌埠市一家未进行牛传染性鼻气管炎及病毒性腹泻免疫的规模化奶牛场的94份奶牛血清样品,分别使用牛传染性鼻气管炎及病毒性腹泻抗体ELISA试剂盒进行检测,共检出牛传染性鼻气管炎阳性血清65份,阳性检出率为69．15％;病毒性腹泻阳性血清83份,阳性检出率为88．30％。结果表明,该规模化奶牛场存在牛传染性鼻气管炎及病毒性腹泻的感染和接触史,应采取净化措施进行控制。  相似文献   

14.
本研究以牛传染性鼻气管炎gE基因的原核表达产物为抗原建立检测IBRV抗体间接ELISA检测方法.通过DANStar比对分析牛传染性鼻气管炎病毒gE蛋白序列,通过亲和层析对重组表达产物进行纯化,经Western blot检测证明重组表达产物能被IBRV标准性鼻气管炎gE蛋白长度为500 bp的特异性肽序列,随后构建了pC...  相似文献   

15.
Efficient methods of diagnosis and prophylaxis of infectious bovine rhinotracheitis must consider the concept of latency of the etiological agent, infectious bovine rhinotracheitis virus (Bovine herpesvirus 1; BHV 1). The identification of BHV 1 in nasal mucus samples or a rise in specific antibodies have to be cautiously interpreted, because they can signify either a primary infection or a reexcretion of the virus after reactivation. The isolated virus can also either be a vaccine or a virulent strain. Another aspect of BHV 1 infection diagnosis is the detection of latent carriers, which are able to transmit the virus to uninfected animals; delayed hypersensitivity test seems to be a good candidate. The classical methods of prophylaxis protect the animal against the disease, but they should also impede the reexcretion of virulent strains by latent carriers. Since, in several countries, attenuated viruses are used as vaccines, a special emphasis has to be laid on the persistence of these vaccine viruses in a latent form in the bovine population.  相似文献   

16.
A single-dilution, sensitive and specific monoclonal antibody-based blocking enzyme-linked immunosorbent assay (B-ELISA) was developed as an alternative to the cumbersome virus neutralization test (VNT) for detection of equine herpesvirus-1 (EHV-1) antibodies. Neutralizing monoclonal antibodies (1H6 and 9C6) raised against EHV-1 (Hisar-90-7 strain) and sera from 70 horses (30 known negative and 40 known positive for EHV-1 antibodies by VNT) were used for standardization of the B-ELISA. Using a single serum dilution of 1:250 in B-ELISA, 100% specificity was obtained with both monoclonal antibodies (Mabs) in comparison to VNT. Similarly, the sensitivity of the B-ELISA was 92.5% and 100% with 1H6 and 9C6 Mabs, respectively. A very high correlation coefficient (r = 0.85) was observed between B-ELISA and VNT that was significant at the p < 0.01 level. B-ELISA detected a more than 3-fold rise in antibody titres in paired serum samples collected from mares aborting owing to EHV-1 infection. Mab 9C6 was chosen for testing 231 field sera from apparently healthy vaccinated and non-vaccinated horses from organized breeding farms belonging to 11 Indian states, and from Bhutan, by B-ELISA and VNT. There was very good agreement between the results obtained by both VNT and B-ELISA (K = 0.9438). Of 231 field sera, 144 samples were negative for EHV- 1 antibodies by both VNT and B-ELISA and 81 were positive by both tests. Two samples negative by VNT were found positive in B-ELISA. On the other hand, four weakly positive samples in VNT (VN antibody titre 0.9 1.2 log10) were negative in B-ELISA. The Mab (9C6)-based B-ELISA was found to be a suitable alternative to VNT for screening large numbers of field sera and enabled confirmatory EHV-1 serodiagnosis.  相似文献   

17.
Isotype-capture ELISAs for BCV-specific IgA and IgM were developed and tested on milk and serum samples from Swedish cattle. The capture ELISAs showed higher sensitivity than indirect ELISAs for detection of BCV-specific IgA and IgM. In the capture ELISAs the agreement between detection in milk and serum samples was 94% for IgA and 86% for IgM. The correlation between log(10) titres in milk and serum was r=0.82 (P<0.001) for IgA and 0.84 (P<0.001) for IgM. Milk seemed a better target than serum for diagnosing specific IgA at low levels. There was no variation in the isotype-specific BCV antibody titres between healthy quarters of the same udder, but subclinical mastitis was associated with higher levels of IgA antibodies and weak false IgM positive reactions in undiluted milk. Bovine IgA and IgM antibodies in milk and serum showed high stability towards freezing and thawing and storage at room temperature.The antibody responses to BCV were followed in milk and serum from six dairy cows and in serum from four calves for a period of 1 year after an outbreak of winter dysentery (WD). In this outbreak some animals became reinfected with BCV. The IgA and IgM capture ELISAs differentiated between primarily BCV infected and reinfected animals. In the primarily infected cattle, IgM antibodies were first detected in milk and serum four to nine days after the first WD symptoms observed, and were subsequently detected for at least 2-3 weeks. IgM was also detected in the reinfected cows, but mostly at lower levels and for a shorter period of time than in the primarily infected animals. In milk, however, the IgM response of the reinfected cows was detected for a longer period of time than in serum. Six months after the outbreak, IgA was still detected in both serum and milk of all six cows and also in serum of one calf. The reinfected cows showed higher and more long-lasting peak levels of IgA in milk and serum than the primarily infected cows, indicating boosting of the IgA response.  相似文献   

18.
Antibody (immunoglobulin G (IgG), IgM or IgA) levels relative to ferritin in six foal sera (three male and three female) after birth (day 0 and 2, 6, 10, 20, 28, 36, 40, 52 and 56 weeks of age) were semi‐quantitatively measured with normalization with antibody activity to ferritin in one adult horse serum. After addition of horse spleen ferritin to the serum sample, the complex formed between antibodies to ferritin in the serum and ferritin was co‐immunoprecipitated using antibody to horse spleen ferritin. Antibody classes of the co‐immnoprecipitate were detected with antibodies specific for horse IgG, IgM or IgA heavy chain. Six adult horse serum samples were found to have ferritin‐binding activities in all immunoglobulin classes examined. Although ferritin antibody activities (IgG, IgM and IgA) were scant in the foal sera before sucking colostrum (day 0), their activities increased at 2 weeks of age. IgG antibodies showed a biphasic response and IgM antibody activity increased up to 40 weeks of age. Antibody (IgG, IgM and IgA) activities to ferritin in three colostrum samples were significantly higher than in adult horse serum samples. These results demonstrate that antibody to ferritin in foal serum is derived from colostrum after birth and is produced thereafter.  相似文献   

19.
More than 400 small ruminant sera from Za?re were screened for antibodies to IBR, CHV2, BVD, bovine and ovine PI3, BRS and rinderpest viruses. Sera from local animals were negative for BVD, PI3 and rinderpest viruses: 8% of sera were positive for IBR virus, all with higher titers to CHV2; 31% of sera were positive to BRS virus.  相似文献   

20.
为建立检测牛传染性鼻气管炎病毒(IBRV)血清抗体的阻断ELISA方法,本研究以经蔗糖密度梯度离心法纯化的IBRV作为免疫原制备1株单克隆抗体(MAb),命名为cp-1-1。经间接ELISA、IFA和western blot鉴定,该MAb与IBRV呈阳性反应,与牛病毒性腹泻病毒(BVDV)及牛副流感病毒3型(BPIV3)呈阴性反应,具有较强的特异性。质谱分析结果显示MAb cp-1-1识别的表位位于IBRV VP8蛋白。以纯化的IBRV作为包被抗原、MAb cp-1-1作为检测抗体,建立检测IBRV血清抗体的阻断ELISA方法。该检测方法的抗原包被量为0.89μg/孔,样品稀释度为12,检测抗体MAb量为1.3μg/孔,二抗稀释度为15000。利用50份IBRV抗体呈弱阳性的牛血清(中和抗体效价为14~116)作为标准参考血清,确定该检测方法的阻断率Cut Off值为52.06%,即阻断率高于52.06%时判为阳性,低于52.06%时判为阴性。阻断ELISA方法特异性试验显示仅IBRV阳性血清检测为阳性,而BVDV、BPIV3、牛腺病毒3型(BADV-3)和O型口蹄疫病毒(O-FMDV)阳性牛血清均检测为阴性,表明该方法具有较强的特异性;该方法可检测的最低中和抗体效价为14,与病毒中和试验的敏感性一致,表明该方法具有较高的敏感性;重复性试验显示该方法批内、批间变异系数均小于10%,显示较好的重复性。对130份现地牛血清检测结果显示,该方法与病毒中和试验的符合率为98.46%。用该方法对某牛场接种IBRV灭活疫苗的牛血清进行检测,抗体阳性率为99.51%(205/206)。另外,采用该方法对我国8个省(市、自治区)的801份牛血清进行检测,IBRV的抗体阳性率为41.6%(333/801)。本研究建立的阻断ELISA方法可以用于IBRV疫苗免疫监测和血清流行病学调查,为我国IBR的防控提供技术支持。  相似文献   

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