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1.
应用云南畜牧兽医研究所研制的蓝舌病病毒:(BTV)羟胺灭活苗接种绵羊178只。接种后42天以琼扩试验检测免疫接种前琼扩阴性羊45例和琼扩阳性羊15例.均无改变。接种后205天攻毒,免疫羊保护7/10,对照羊蓝舌病阳性数为8/10.  相似文献   

2.
用IBD复合油佐剂组织灭活苗(简称IBD油苗)接种14日龄蛋雏鸡(0.2ml/只)后7、14日用琼扩试验(AGP)测IBD抗体,全部阴性。于32日龄第2次接种上述IBD油苗(1 ml/只)后26、46天测IBD抗体,结果全部阳性。平均效价为1.31g_3及3.5 Ig_2.雏鸡于第1次注IBD油苗后18天(32日龄)和第2次注苗后90天(122日龄)用IBD强毒攻击,均获得全数保护。于现地用IBD油苗接种500只青年蛋鸡(0.5ml/只),注苗后第6天,同舍来注苗的3批青年蛋鸡(499只)暴发IBD,6天内死亡率达7.6~53.6%,而注苗鸡全获保护。  相似文献   

3.
一、肉鸡上试验及应用 选用某禽蛋公司92年3月18日自孵的AA商品苗鸡10800只,饲养在本场鸡舍。试验分为二组,①试验组:10728只雏鸡于21、36日龄各注射自制IBD卵黄抗体(琼扩效价1:64以上)0.5毫升进行被动免疫。②对照组:72只雏鸡于18日龄用进口D_(78)苗免疫,疫苗由郊区畜牧兽医站提供,批号:910927。两组均于首免后7天、14天用琼扩试验监测。其他饲养管理均相同。 试验组在整个饲养期内未发生IBD,对照组于25日龄发生IBD,一周内共死亡12只,死亡率16.6%,琼扩监测结果均为阴性。  相似文献   

4.
应用仔猪副伤寒琼扩抗原,对副伤寒高兔猪进行检测,第一次注射后第5天3/3阳转,抗体持续34-72天;对副伤寒弱毒苗免疫猪进行琼扩检测,于注苗后7天9/11阳转,14天11/11阳转,抗体持续7-40天,该法与猪布氏杆菌病、猪巴氏杆菌病、猪丹毒、猪弓形体病、猪瘟、猪密蠓旋体病、猪链球菌病均无交叉反应。用本法检测1288份现地血清,阳性率3.26%,试验肜该法监测仔猪副伤寒特异性强,操作简便、快速、具  相似文献   

5.
以禽呼肠病毒S1133毒株尿囊腔接种10日龄无特定病原体(SPF)鸡胚,无菌收集24 h~120 h的死胚尿囊液,加1 mL/L的甲醛灭活,浓缩制得琼扩抗原;用该抗原制成灭活苗两次免疫SPF鸡,无菌采血分离制备阳性血清;选用SPF鸡无菌采血分离制备阴性血清.经过效价测定制备16单位琼扩抗原,8单位阳性血清;取免后的SPF鸡血样4个20~2-5稀释分别与1单位~8单位琼扩抗原做琼扩试验,确定了应该用4单位或8单位琼扩抗原做工作抗原;并以血清中和试验为参照证明琼扩试验在血清抗体监测中的可行性.本研究成功制备了禽呼肠病毒琼扩抗原与血清,为用琼扩试验定量检测禽呼肠病毒感染禽和疫苗免疫禽的抗体奠定了基础.  相似文献   

6.
本试验建立了检测鸡病毒性关节炎病毒的ELISA双抗体夹心法.取代反应、阻断试验均为阴性,与NDV、MDV和IBDV无交叉反应,对已知阳性标本的检测均为阳性;其敏感性比琼扩试验高80倍以上,这表明ELISA夹心法具有较高的特异性和敏感性.在人工感染后2~27d,关节滑膜、腱鞘和脾脏中病毒检出率为100%;还从肝脏、法氏囊和脑组织中检测到病毒.  相似文献   

7.
应用HRP—SPA—ELISA检测猪瘟病毒抗体的试验研究   总被引:2,自引:0,他引:2  
应用猪瘟兔化弱毒PEG沉淀抗原、HRP—SPA建立了HRP—SPA—ELISA检测猪瘟病毒抗体的方法。通过对5份阴性血清和32头仔猪注苗前及注苗后不同时期256份系列血清的检测以及阻断试验,证明本法具有较高的敏感性和特异性。猪瘟抗体ELISA效价与兔体中和试验效价比较,二者呈显著的正相关(r=0.8072)。对32头仔猪系列血清的检测结果表明,免疫母猪(配种前45d左右注苗)所生的30~40日龄未注苗仔猪的母源抗体水平不很高,有1/3在阴性范围内,注苗后15d,ELISA抗体开始上升,90d达峰值;90~120d大部分猪的抗体水平迅速下跌到略高于60d时的抗体水平上,保持稳定约2个月左右,但也有一少部分猪90d后抗体水平下跌缓慢,一直保持相当高的水平。  相似文献   

8.
IBD试纸条对中强毒力苗毒及分离野毒的检测研究   总被引:1,自引:0,他引:1  
以琼扩为对照,应用IBD试纸条分别检测了人工感染鸡、疫苗接种鸡、自然发病鸡3种不同类型的法氏囊样品以及不同的疫苗。毒株测定结果显示,IBD苗毒228E、MB、V877、NF8、QC试纸条均为阳性,琼扩对照均为阴性,分离强毒H4、He3、Y2试纸条及琼扩对照均为阳性。法氏囊样品测定结果显示,自然感染病例,H4、He3、Y2感染鸡,228E、MB、V877、NF8接种鸡两种方法测定均为阳性,QC则为阴性。对法氏囊样品的检测结果证明,IBD试纸条具有特异、敏感、快速的特点,其敏感性高于琼扩的敏感性,为IBD诊断提供了新方法。  相似文献   

9.
本试验测定了马传贫弱毒疫苗免疫马与传贫病马琼脂扩散沉淀抗体(沉淀抗体)及补体结合反应抗体(补反抗体)活性的耐热曲线。证明免疫马的抗体较传贫病马的耐热性差。64℃水浴15分钟后,免疫马血清沉淀抗体完全丧失反应活性,而传贫病马血清一般多能保持阳性反应。用65℃水浴15分钟处理琼扩阳性反应血清,结果传贫病马血清142份中的128份仍为琼扩阳性,传贫病马注苗后的血清5份均呈琼扩阳性;免疫马血清37份则全部转为阴性反应。65℃水浴15分钟后,免疫马血清补反抗体完全丧失反应活性,而传贫病马血清仍可呈阳性反应。用65℃水浴15分钟处理补反阳性血清,结果传贫病马血清40份、强毒接种马血清2份的补反仍保持阳性;免疫马血清42份则均转为阴性。  相似文献   

10.
某单位养殖的羊群在接受当地兽医站羊痘防疫时,引发急性巴氏杆菌病。在当日注苗的9户中就有7户羊发病,于注苗后10h先后出现临床症状。共注苗267只,发病35只,死亡23只(其中1.5~3月龄羔羊20只,瘦母羊3只)。发病率13%,病死率65.7%。  相似文献   

11.
A maedi-visna virus CA-TM fusion protein ELISA (MVV ELISA) was evaluated for the detection of antibody in sheep infected with North American ovine lentivirus (OvLV). The results of the MVV ELISA were compared with other assays for OvLV antibody and with viral infection in an intensively studied group of 38 sheep with a high prevalence of OvLV infection and disease. The sensitivity, specificity, and concordance of assays for OvLV antibody (MVV ELISA, indirect ELISA, Western blot, and AGID), virus (virus isolation, PCR, antigen ELISA), and OvLV-induced disease in each animal were compared with OvLV infection status as defined by a positive result in two or more of the assays. Five sheep met the criteria for absence of OvLV infection. The sensitivity of the MVV ELISA in detecting OvLV infected sheep was 64%, whereas the sensitivity of the other three tests for antibody ranged from 85 to 94%. All the antibody assays were 100% specific in this group of animals. Of the assays for virus, the PCR test had the highest sensitivity and the best concordance with OvLV infection, but it also had the lowest specificity of any of the virus or antibody assays. Among the antibody tests, the concordance of the MVV ELISA compared most favorably with the AGID test for detecting OvLV-infected sheep. Analysis of serum samples from 28 lambs experimentally-infected with one of three North American strains of OvLV suggested that there were no significant strain differences detectable by antibody assay. Twenty virus-inoculated lambs were positive by both the MVV ELISA and the AGID test, five lambs were MVV ELISA negative and AGID test positive, and three lambs were MVV ELISA positive and AGID test negative. No pre-inoculation samples were positive by either assay. In a longitudinal study involving seven lambs, antibodies to OvLV were detected by AGID 3-5 weeks post-inoculation, but were not detected by MVV ELISA until 5-10 weeks post-inoculation. Among 128 naturally and experimentally-infected sheep that were seropositive in the AGID test, the overall sensitivity of the MVV ELISA was higher in the naturally infected sheep (84%) than in the experimentally infected sheep (69%). The data indicated that the MVV ELISA represents a less sensitive, but specific alternative for the detection of OvLV antibodies.  相似文献   

12.
The production of interferon-gamma (IFN-gamma) in response to Johnin purified protein derivate was measured in samples of the prescapular lymph node (PLN) from 10 sheep, aged 2 years, and nine sheep, aged 1 year that had been inoculated orally with Mycobacterium avium subsp. paratuberculosis within their first month of life. Ten non-inoculated sheep, aged 1 year, constituted the negative control group. The results obtained in the PLN IFN-gamma assay were compared with those derived from serological tests: a complement fixation test (CFT), agar gel diffusion test (AGID) and enzyme-linked immunosorbent assay (ELISA), as well as an IFN-gamma test on samples of blood. Among the 19 inoculated sheep, 16 gave positive reactions in the PLN IFN-gamma assay on samples incubated overnight, and 18 tested positive when the assay was applied to PLN samples incubated for 48h. In comparison, three, four and seven inoculated sheep gave positive reactions in the ELISA, CFT and in the blood IFN-gamma assay on samples incubated overnight, respectively. The AGID and IFN-gamma assay on blood samples incubated for 48h detected eight inoculated animals. Twelve inoculated sheep, that tested positive in the PLN IFN-gamma assay were clinically normal, gave negative results in an IS900-based polymerase chain reaction (PCR) assay on samples of ileum and ileocaecal lymph node and had no histological evidence of paratuberculosis, but tested positive on more than two occasions in sequential serological testing before necropsy. None of the 10 non-inoculated sheep tested positive in the AGID, CFT, ELISA, blood IFN-gamma assay on samples incubated overnight and for 48h or the PLN IFN-gamma assay on samples incubated overnight, but one gave a positive result in the PLN IFN-gamma assay on samples stimulated for 48h. It is likely that the positive reactions obtained by the PLN IFN-gamma assay in the 12 inoculated sheep that tested negative in the PCR assay and histopathological examination represents immunological evidence of latent infection or previous exposure to M. paratuberculosis rather than active infection.  相似文献   

13.
The present study was designed to evaluate a commercial ELISA kit (Institut Pourquier) for the diagnosis of ovine and caprine paratuberculosis under Australian conditions and to compare its accuracy with the existing AGID test. The sensitivity of the ELISA in sheep and goats was 34.9% and 56.4%, with a specificity of 98.8% and 100.0%, respectively. Sensitivity of AGID was 13.8% for sheep and 39.5% for goats, with specificity of 100.0% for both species. The sensitivity of the ELISA in sheep depended on the category of histological lesions. AGID and ELISA were conditionally independent, and appeared to detect overlapping but distinct subgroups of infected animals. The ELISA was significantly more sensitive than the AGID. The ELISA was simple to perform, robust and repeatable. Coefficients of variation of <12.0% were observed for positive and negative controls included on 193 plates over a 10-month period and there was a high level of intraassay repeatability with 12.0% of the duplicate samples having CV of >15.0%.  相似文献   

14.
The sensitivities and specificities of an absorbed enzyme-linked immunosorbent assay (ELISA) and an agar-gel immuno-diffusion (AGID) test for the detection of Johne’s disease in sheep were estimated using data from six known infected and 12 assumed uninfected sheep flocks. Sensitivities were estimated for all histologically positive sheep, as well as by histological lesion score, lesion type (paucibacillary or multibacillary) and sheep body-condition score, with ELISA sensitivities estimated at 95 and 99% specificity. Logistic-regression analysis was used to test for significant effects of lesion score and condition score, with flock included in the model as a random effect.

Estimated specificities were 95% (95% CI: 93.4, 95.6%) and 99% (98.4, 99.4%) for ELISA cut-point ratios of 2.4 and 3.6, respectively, and 100% (99.7, 100.0%) for the AGID. Estimated sensitivities for all infected sheep were 41.5% (35.0, 48.3%), 21.9% (16.6, 27.9%) and 24.6% (19.1, 30.7%) for ELISA cut-point ratios of 2.4 and 3.6 and for AGID, respectively. Sensitivities of all tests and cut-points varied significantly between flocks and between categories of lesion score and condition score. Sensitivity ranged from 25 to 73, 10 to 47 and 9.2 to 63% between flocks, for the ELISA with cut-points of 2.4 and 3.6, and for the AGID, respectively. Sensitivity was highest in thin sheep and in sheep with multibacillary lesions. The effects of lesion type and condition score on test sensitivity were significant in the logistic regressions for the AGID and ELISA at both cut-points and the flock effect was significant for the AGID but not for the ELISA at either cut-point.  相似文献   


15.
Our objective was to estimate the sensitivity and specificity of the agar-gel-immunodiffusion test (AGID), the ELISA, and the skin test for the detection of Mycobacterium avium subspecies paratuberculosis (MAP) in sheep using Bayesian methods without a gold standard. Fourteen flocks (2 465 sheep) were used. Five flocks (450 sheep) were considered MAP non-infected and 9 flocks (2 015 sheep) had sheep infected with MAP. Sheep were skin tested and blood was collected for AGID and ELISA testing. Results were analyzed using a Bayesian 3-test in 1-population model fitted in WinBUGS. The model allowed for dependence (correlation) between the two serologic tests, but these two tests were assumed to be conditionally independent of the skin test. The estimated specificity was 99.5% (95% PI of 98.9-99.9%) for the AGID; 99.3% (98.4-99.8%) for the ELISA using an optical density measured cutoff of 0.20; 99.2% (98.1-99.8%) using a cutoff of 0.15; 97.5% (95.8-98.7%) using a cutoff of 0.10; and 98.7% (97.3-99.5%) for the skin test. The estimated sensitivities were 8.3% (6.2-10.7%) for the AGID; 8.0% (6.0-10.4%), 10.6% (8.3-13.1%), and 16.3% (13.5-19.4%) for the ELISA using the cutoffs 0.20, 0.15, and 0.10 respectively; and 73.3% (62.3-85.8%) for the skin test. The skin test was specific in non-infected populations and sensitive in infected populations, although in some cases a positive skin test might represent MAP exposure rather than infection. The AGID and ELISA were specific but lacked sensitivity. The AGID and ELISA consistently identified two different populations of infected sheep with only moderate overlap between positive test results.  相似文献   

16.
用昆虫杆状病毒表达系统获得的马传染性贫血病病毒( E I A V) 核心蛋白( Gag) 和 P26 蛋白, 作为免疫琼脂双扩散( A G I D) 和酶联免疫吸附试验( E L I S A) 抗原。对76 份已知马传贫非特异性血清进行检查, 同时与市售 A G I D 和 E L I S A 试剂盒作比较。证明, 用表达蛋白作抗原的 A G I D 和 E L I S A 检测结果均为阴性反应, 而用市售 A G I D 试剂盒检查有54 份马血清出现非特异性反应, 市售 E L I S A 试剂盒检查也出现了非特异性反应, O D 值比表达抗原 E L I S A 高35 倍。初步证明在 A G I D 和 E L I S A 法中, 表达抗原优于常规马传贫病毒抗原。  相似文献   

17.
对选自内蒙古4个旗(县)的213只羊采用通过琼脂扩散试验(AGID)、皮内变态反应(DTH)、补体结合反应(CF)和酶联免疫吸附试验(ELISA)4种检测方法进行3次羊副结核病检测,以比较4种检测方法的检出率。结果表明,琼脂扩散试验、皮内变态反应、补体结合反应和酶联免疫吸附试验的检出率分别为3.8%、4.7%、5.2%和11.3%。其中以酶联免疫吸附试验的检出率最高,并且该方法简便、灵敏,具有实际应用和推广价值,对内蒙古地区羊副结核病检测方法的选择有一定的指导意义。  相似文献   

18.
To investigate Brucella infection in cattle, sheep, goat, reindeer and yak in Mongolia, serological reactions of Brucella-infected and -vaccinated domestic animals were compared by the agar gel immunodiffusion (AGID) test with a polysaccharide (poly-B) of the B. Abortus strain S-19. The sensitivity and specificity were compared with conventional serological tests that are commonly used in Mongolia, such as the rose Bengal test, the tube agglutination test and the compliment fixation test. A total of 73.3, 100, 100, 95.8 and 61.9% of the sera from suspected cattle, yak, goat, sheep and reindeer, respectively, that were positive in the compliment fixation test, were also positive in the AGID test. Sera from vaccinated cattle, sheep and goat were positive over 90% by conventional tests 3 months after vaccination, but were negative by the AGID. These results suggest that the AGID test may be useful to differentiate infected and vaccinated animals in the field.  相似文献   

19.
An enzyme-linked immunosorbent assay (ELISA) test was developed for the detection of specific antibodies against the unique infectious anemia (EIA) virus in equine sera. The ELISA test was faster and more sensitive when compared with the classic test of agar gel immunodiffusion (AGID). A total of 200 sera were tested: 100 from negative horses and 100 from positive horses by AGID. The ELISA test showed 92 horse sera negative and 100 horse sera positive by AGID with values of optical density (OD) less than 0.139 and higher than 0.139, respectively. Eight horse sera were negative by AGID and higher than 0.139 by ELISA. Six of these became AGID positive also when re-tested 30 days later, and two were of the horses that showed clinical signs of EIA and died before re-testing.  相似文献   

20.
OBJECTIVE: To determine the sensitivity and specificity of an absorbed ELISA and an AGID test for the detection of clinical and subclinical paratuberculosis in sheep. DESIGN: By testing a panel of sera from 1257 Australian Merino and crossbred sheep greater than 1 year of age, of which 1137 sheep were not infected with Mycobacterium avium subsp paratuberculosis and 120 sheep had paratuberculosis. PROCEDURE: Sera were collected from 457 sheep in Victoria and 800 sheep in Western Australia. Presence of M a paratuberculosis infection in Victorian sheep was determined by histological examination of intestinal tissues, whereas sheep from Western Australia were presumed to be free of Johne's disease. The ability of an absorbed ELISA to discriminate between infected and uninfected sheep was described by test sensitivity and specificity, the distribution of ELISA OD, and the area under a receiver operating characteristic curve. RESULTS: The absorbed ELISA had a specificity of 98.2 to 99.5% (CI) and a sensitivity of 35 to 54% (CI). In sheep from infected flocks in Victoria, the AGID test had a specificity of 99 to 100% (CI) and a sensitivity of 38 to 56% (CI). The sensitivity of serological tests was higher in sheep with a body condition representative of the lower quintile of their flock of origin. CONCLUSION: The AGID test and absorbed ELISA are useful tests for the detection of ovine paratuberculosis. Although the tests had a similar accuracy, they detected different subpopulations of infected sheep with only moderate overlap. The AGID test had a higher specificity than the absorbed ELISA.  相似文献   

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