首页 | 本学科首页   官方微博 | 高级检索  
相似文献
 共查询到20条相似文献，搜索用时 484 毫秒
1.
Serially collected epithelial samples from lesions in the mouth and on the feet of calves experimentally infected with foot-and-mouth disease (FMD) type O1 BFS 1860 were assayed for the presence of FMD viral antigen using a double antibody sandwich enzyme-linked immunosorbent assay (ELISA) and a complement fixation (CF) test. The amount of infectious virus in each sample was also determined. FMD viral antigen was detected by ELISA in 70 per cent of the mouth samples and 92 per cent of samples from the feet. The CF test was less sensitive; it detected antigen in 44 per cent of mouth and 85 per cent of foot samples. In mouth samples the amount of antigen decreased rapidly becoming undetectable by the fourth day of sampling whereas in foot samples the quantity of antigen declined more slowly, and could be detected until the seventh day of sampling. Therefore it was concluded that the age of lesion and the site from which epithelial samples are collected are both important determinants in the laboratory diagnosis of FMD. In cattle, foot lesions are more likely than mouth lesions to yield antigen and to remain positive for a longer period.  相似文献   

2.
OBJECTIVE: To evaluate and implement rapid molecular diagnostic techniques for the detection of foot and mouth disease virus (FMDV) suitable for use in Australia. DESIGN: Two PCR TaqMan assays targeted to the FMDV internal ribosome entry site or the 3D polymerase coding region for the rapid detection of FMDV were evaluated using non-infectious materials to determine the test most appropriate for implementation as part of Australia's national preparedness for the rapid detection and diagnosis of FMD outbreaks. RESULTS: Two published tests (PCR TaqMan assays targeted to the FMDV IRES region or the FMDV 3D polymerase coding region) were evaluated for their ability to detect FMDV genetic material in non-infectious FMDV ELISA antigen stocks held at Australian Animal Health Laboratory. Both tests were able to detect FMDV genetic material from strains O1 Manisa, O-3039, A22, A24, A Malaysia, C, Asia 1 and SAT 1, 2 and 3. With the exception of Asia 1, the TaqMan assay targeted to the FMD 3D polymerase coding region had Ct values equal to or lower than for the TaqMan assay targeted to the IRES region suggesting that this test may provide broader serotype detection and sensitivity. However, the TaqMan assay directed to the FMDV IRES is the only one to date to have undergone substantial evaluation using clinical samples collected during an outbreak. The greatest differences observed were for O-3039, SAT 1, and 3. CONCLUSION: Given the ease of setting up both tests, AAHL currently runs both tests on highly suspect FMD investigations to provide independent confirmation of the absence of FMDV because the tests are focused on two independent regions of the FMDV genome. These tests add substantially to Australia's preparedness for FMD diagnosis complementing the already well-established virus isolation and antigen capture ELISA tests for index case diagnosis of FMD in Australia.  相似文献   

3.
在2019年春、秋季集中免疫后,分别采集四川省21个市（州）被抽检县（区）的规模场、散养户、屠宰厂的猪血清共2 306份,采用ELISA方法进行口蹄疫O型免疫抗体检测。结果显示,2019年全省猪O型口蹄疫免疫抗体的平均合格率为85.0%（1 960/2 306）,其中春季检测1 556份,合格1 370份,合格率为88.05%;秋季检测750份,合格590份,合格率为78.67%。结果表明我省2019年春秋两季猪O型口蹄疫疫苗免疫均取得理想效果,能够对猪群提供免疫保护。  相似文献   

4.
作者以固相法合成特异性FMDV主要保护性抗原VP1上的表位肽,将其与载体蛋白BSA偶联,作为包被抗原,制备检测抗O型口蹄疫病毒(FMDV)抗体的ELISA试剂盒,并对该试剂盒进行方法考核.结果表明该方法的敏感性为95.12%,特异性为100%.检测199份血清标本,与UBI FMD VP1试剂盒的符合率达到98.49%,与液相阻断ELISA试剂盒的符合率达到96.98%.该多肽ELISA试剂盒特异、敏感、稳定、操作简便,可用来监控口蹄疫抗体水平.  相似文献   

5.
为评价口蹄疫病毒A型竞争ELISA(cELISA)抗体检测试剂盒在流行病学调查中的应用前景,对2017年从福建省三明市采集的336份黄牛、奶牛、羊和猪血清样品,用A型cELISA抗体检测试剂盒进行抗体检测。结果显示,92份黄牛血清、92份羊血清、92份猪血清、60份奶牛血清的A型抗体阳性率分别为13.04%、11.96%、20.65%、86.67%。从上述4种血清中,各挑选10份血清(阴性、阳性各5份)共40份,采用口蹄疫病毒液相阻断ELISA(LPB-ELISA)抗体检测试剂盒进行验证。结果显示:cELISA检测为阳性的20份血清中,用LPB-ELISA检出阳性19份;cELISA检测为阴性的20份血清中,用LPB-ELISA检出阴性17份;两种方法的κ值为0.8,总符合率为90.00%。结果表明,A型cELISA试剂盒与LPB-ELISA试剂盒的符合率和一致性均较高,可用于口蹄疫流行病学调查和血清学监测。  相似文献   

6.
The aim of this study was to evaluate the value of commercially available kits for the detection of foot-and-mouth disease (FMD) virus infection in vaccinated cattle. The cattle were vaccinated with a commercial aqueous FMD vaccine type A24 and subsequently challenged 28 days post vaccination with homologous FMD virus. Seven of eight animals were protected from clinical disease and all became carriers. They were bled sequentially for up to 130 days post infection and samples of sera were tested with three ELISA kits: CHEKIT FMD-3ABC, Ceditest FMDV-NS and SVANOIR FMDV 3ABC-Ab ELISA. The Ceditest kit appears to be relatively higher sensitive than the others. When examined with this ELISA, all cattle developed of FMDV nonstructural proteins (NSPs) antibodies and remained positive throughout the period of the experiment. The response of antibodies against 3ABC antigen delayed in two cattle challenged with FMDV A24 virus. One of the cattle reacted negatively in Svanoir ELISA kit and sera from two animals were found negative in CHEKIT ELISA. It can be concluded that all tested kits can be a promising tool for FMD control and eradication campaigns in situation where emergency vaccination was applied.  相似文献   

7.
During 1999, 11 outbreaks of foot and mouth disease (FMD) were declared in the east and central part of Morocco. All the FMD clinical cases reported were cattle. In order to analyse the serological status of sheep from the FMD outbreak areas, 598 sheep sera were tested using a liquid-phase blocking ELISA (LPBE) to detect antibodies against FMDV structural proteins. The study confirmed the presence of FMDV specific antibodies in 77 clinically normal sheep, indicating that unrecognised FMDV-infected sheep could represent a potential risk of FMD dissemination in Morocco.Subsequently, sera from flocks of sheep that had been exposed to FMD outbreaks were assayed by an indirect ELISA using the recombinant FMDV non-structural protein 3ABC expressed in E. coli to evaluate the potential use of this serological test in future epidemiological studies and the development of FMD control strategies. The results indicated that the 3ABC-ELISA was able to detect antibodies indicative of infection with FMDV in asymptomatic sheep in field conditions.  相似文献   

8.
This study was conducted to determine if humoral antibody response of foot-and-mouth disease (FMD) vaccine improved in 8-week-old growing pigs born to well-vaccinated sows pre-treated with 60 mg of poly-γ-glutamic acid (γ-PGA) three days before vaccination. Antibody against FMD virus serotype O was measured 0, 2, 4 and 6 weeks post-vaccination, using a PrioCHECK FMDV type O ELISA kit. The results showed that positive antibody reactions against FMDV serotype O antigen among a component of the vaccine significantly increased in response to pre-injection with γ-PGA.  相似文献   

9.
使用牛用口蹄疫AsiaI-O型双价灭活苗与猪用口蹄疫O型灭活苗分别接种50d商品猪和怀孕90d种猪,免疫前和免疫后的3w及7w进行抗体水平检测。O型口蹄疫采用正向间接血凝试验、AsiaI型口蹄疫采用液相阻断ELISA检测。同时对接种猪进行免疫应激观察。结果显示：猪使用牛用口蹄疫AsiaI—O型双价灭活苗3w后口蹄疫AsiaI的抗体水平十分低,最高只有5％,口蹄疫O型抗体合格率在35％以上;而注射猪用口蹄疫O型灭活苗的O型抗体水平合格率只有35％。而牛用口蹄疫AsiaI-O型双价灭活苗两次接种后,AsiaI和O型抗体水平均达到大于70％的要求。  相似文献   

10.
本研究旨在表达口蹄疫病毒(FMDV)的VP1全基因并制备特异性的多克隆抗体。利用PCR方法扩增Asia 1 IND 49197株VP1全基因,将其克隆至原核表达载体pET-30a(+)中,在大肠杆菌BL21中进行表达。SDS-PAGE结果显示表达产物分子量约为31.6ku,以包涵体的形式存在。通过Ni-NTA Purification System纯化后进行western blot和间接ELISA分析,结果显示重组蛋白能够被FMD阳性血清识别,具有良好的反应性。将纯化的重组蛋白免疫新西兰白兔制备多克隆抗体,ELISA测定抗体效价为1∶20480,病毒中和试验测定抗体效价为1∶64。本研究所表达的VP1蛋白可用于开发检测Asia1口蹄疫抗体的诊断试剂,所制备的多克隆抗体为进一步研究VP1的结构、功能以及抗原表位的鉴定提供了条件。  相似文献   

11.
Lu Z  Cao Y  Guo J  Qi S  Li D  Zhang Q  Ma J  Chang H  Liu Z  Liu X  Xie Q 《Veterinary microbiology》2007,125(1-2):157-169
Non-structural protein (NSP) 3ABC antibody is considered to be the most reliable indicator of present or past infection with foot-and-mouth disease virus (FMDV) in vaccinated animals. An indirect ELISA was established, using purified His-tagged 3ABC fusion protein as antigen, for detection of the antibody response to FMDV NSP 3ABC in different animal species. The method was validated by simultaneous detection of the early antibody responses to NSP and structural protein (SP) in FMDV Asia 1 infected animals. The performance of the method was also validated by detection of antibody in reference sera from the FMD World Reference Laboratory (WRL) in Pirbright, UK, and comparison with two commercial NSP ELISA kits. The results showed that the antibody response to SP developed more quickly than that to NSP 3ABC in FMDV infected animals. In contact-infected cattle, the antibody response to NSP 3ABC was significantly delayed compared with that to SP antibody. The early antibody responses to SP and NSP 3ABC in FMDV inoculated cattle and contact-infected or inoculated sheep and pigs were generally consistent. In pigs, 3ABC antibody was linked to the presence of clinical signs; however, in sheep, subclinical infection was detected by the development of 3ABC antibodies. Therefore, the antibody responses to 3ABC varied between host species. Eight out of 10 positive serum samples from FMD WRL were tested to be positive at cutoff value of 0.2. The rate of agreement with the ceditest FMDV-NS and the UBI NSP ELISA were 98.05% (302/308) and 93.2% (287/308), respectively. The prevalence of 3ABC antibodies reached 71.4% in some diseased cattle herds. The further work is required to evaluation the performance of this method in different animal species and different field situations.  相似文献   

12.
The epidemiological analysis of the cattle population during the eradication plan of foot and mouth disease (FMD) in Argentina clearly indicated a higher incidence of the disease in animals within their first year of age. It is important to improve the efficacy of the vaccination in those animals. In a previous report, we have shown the effect of an immunomodulator, Avridine (Avr), in the enhancement of the immune response elicited by FMD virus (FMDV) vaccines in experimental hosts [Berinstein, A., Pérez Filgueira, M., Schudel, A., Zamorano, P., Borca, M., Sadir, A.M., 1993. Avridine and LPS from Brucella ovis: effect on the memory induced by foot-and-mouth disease virus vaccination in mice. Vaccine 11, 1295-1301]. In this report, we analyze the effect of Avr in the improvement of the anti-FMDV immune response elicited in young animals immunized with a tetravalent vaccine. The anti-FMDV antibody response was evaluated using a liquid-phase blocking sandwich ELISA (LPBE) [Smitsaart, E.N., Zanelli, M., Rivera, I., Fondevila, N., Compaired, D., Maradei, E., Bianchi, T., O'Donnell, V., Schudel, A.A., 1998. Assessment using ELISA of the herd immunity levels induced in cattle by foot and mouth disease oil vaccines. Prev. Vet. Med 33, 283-296] while the cellular response was detected using an antigen specific lymphoproliferative test [Zamorano, P., Wigdorovitz, A., Chaher, M., Fernández, F., Sadir, A., Borca, M., 1994. Localization of B and T cell epitopes on a synthetic peptide containing the major immunogenic site of FMDV O1 Campos. Virology 201, 383-387]. The results show that, while no differences were detected in the cellular response, the anti-FMDV antibody reaction was significantly (<0.05) higher in animals immunized with the immunogen containing Avr. At 90 days post vaccination, 89-100% of the animals immunized with Avr presented predicted protection (PP) higher than 82% while just 50-61% of the animals immunized with vaccine without immunomodulator presented that characteristic. Also, it is shown that the increase in the anti-FMDV antibody titre in animals immunized with the vaccine containing Avr was mediated by an increase in the levels of both IgG1 and IgG2 which presented a significative correlation with LPELISA antibodies titres. It is concluded that the addition of Avr in the FMDV vaccines improve the immune status of the calves, the cattle population that suffers the highest epidemiological risk.  相似文献   

13.
A novel proximity ligation assay (PLA) using a pan-serotype reactive monoclonal antibody was developed and evaluated for the detection of foot-and-mouth disease virus (FMDV) in clinical samples collected from field cases of disease. The FMDV-specific PLA was found to be 100 times more sensitive for virus detection than the commonly used antigen capture-ELISA (AgELISA). As few as five TCID50 were detected in individual assays, which was comparable with the analytical sensitivity of real-time RT-PCR. Although this assay was capable of detecting diverse isolates from all seven FMDV serotypes, the diagnostic sensitivity of the PLA assay was lower than real-time RT-PCR mainly due to a failure to detect some SAT 1, SAT 2 and SAT 3 FMDV strains. In conclusion, this new PLA format has high analytical sensitivity for the detection of FMDV in clinical samples and may prove valuable as a rapid and simple tool for use in FMD diagnosis.  相似文献   

14.
  相似文献   

15.
Foot and mouth disease (FMD) was detected in a wild boar in Southeastern Bulgaria in December 2010. The occurrence and spread of the disease in wild cloven-hoofed animals may pose an unexpected and significant threat to FMD virus (FMDV)-free areas within and outside the European Union. So far, only one well documented experimental infection with FMD in wild boar has been published. In order to obtain more epidemiologically relevant data regarding the disease in wild boar we conducted an experiment with the 2010 Bulgarian FMDV type O isolate. Two young wild boar were challenged while two domestic pigs and two additional wild boar served as contact controls. While the domestic pigs developed severe clinical signs of FMD, the wild boar showed relatively mild course of the disease. Viremia started in contact wild boar 2 days post exposure (DPE) and lasted until 6 DPE. The virus shedding lasted until 9 DPE. On 27 DPE, when the animals were slaughtered, viral RNA was detected in lymphoid tissues and oropharyngeal fluid but no virus could be isolated. Commercial ELISAs and virus neutralisation tests detected antibodies against FMDV on 8 or 6 DPE, respectively. The data of the present study will help to understand FMD in wild boar populations and can be used in models to evaluate the potential role of wild boar in FMD epidemiology.  相似文献   

16.
A rapid double sandwich enzyme-linked immunosorbent assay (ELISA) has been used for the identification and type differentiation of foot-and-mouth disease (FMD) viruses in epithelial tissue samples submitted for diagnosis from the field. No difficulty was experienced in the direct typing of freshly harvested epithelium from recently ruptured vesicles by the complement fixation (CF) test or ELISA. The ELISA was more sensitive and specific, but proved no more efficient than the traditional CF test in the direct typing of samples of poorer quality from many countries overseas where communications are often difficult. However, when both tests were used concurrently, FMD virus typings were confirmed in 27 more samples. Some possible reasons for the failure of ELISA to detect virus in certain cases are discussed.  相似文献   

17.
  相似文献   

18.
用口蹄疫O型固相竞争ELISA检测血清1 150份,口蹄疫O型免疫血清820份,敏感性为91.83%;试剂盒特异性评价检测血清180份,其特异性为89.4%;固相竞争ELISA和液相阻断ELISA共同检测血清100份,其相关性分析为0.964 1,R2为0.929 5,属于高度相关。3批试剂盒检测血清样品50份,检测结果差异不显著,口蹄疫O型固相竞争ELISA抗体检测试剂盒检测结果比较稳定,具有良好的批间可重复性和稳定性。  相似文献   

19.
An indirect ELISA for detecting the IgA antibody against porcine foot-and-mouth disease virus (FMDV) type A was developed by using purified FMDV structural protein VP1 as coating antigen, mouse anti-pig IgA monoclonal antibody as second antibody and HRP-conjugated goat anti-mouse IgG as third antibody.The concentration of coating antigen was optimized as 3.50 μg/mL,the dilution and reaction time of second antibody and third antibody were optimized as 1:10 000 and 30 min, respectively.There was no cross-reactivity with anti-CSFV, PRRSV and other pathogen specific IgA antibodies.The positive detection rate of FMDV type A infectedsamples was above 90%.The coefficient variation of intra-and inter-assay was ranged from 3.16% to 9.76%.The ELISA method described in this study was proved to be specific and rapid for the detection of FMDV specific IgA antibody.It was potential to be applied for detection the level of FMDV specific IgA and evaluate the effect of mucosal immunity.Besides,it provided a new method for clinical diagnosis of foot-and-mouth disease.  相似文献   

20.
  相似文献   

设为首页 | 免责声明 | 关于勤云 | 加入收藏

Copyright©北京勤云科技发展有限公司  京ICP备09084417号