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1.
为了解血浆、血清对布氏杆菌病虎红平板凝集试验检测结果是否有影响,试验取20只成年羊的颈静脉血,分别进行了虎红平板凝集试验和试管凝集试验,结果发现:血清样品布氏杆菌病RBPT、SAT的检测结果均为阴性;血浆样品布氏杆菌病RBPT的检测结果均为阳性,SAT的检测结果均为阴性。  相似文献   

2.
选用国标等同阳性血清,对虎红抗原进行质控,经三次重复试验,最终选定稀释度为1/45时反应阳性,1/55时反应阴性。ELISA试剂盒质控选择国标等同阳性血清,稀释度为1/16时反应阳性,1/64时反应阴性。利用虎红平板凝集试验(RBT)、试管凝集试验(SAT)和抗体竞争ELISA试验三种方法,对甘肃省河西地区4个奶牛场共计1167份奶牛血清进行了检测,结果表明:三种试验方法的符合率分别为:抗体竞争ELISA试验与虎红平板凝集试验符合率为52.7%,与试管凝集试验符合率为34%。虎红平板凝集试验与试管凝集试验符合率为72.1%。抗体竞争ELISA试验方法最为敏感。  相似文献   

3.
应用平板凝集和试管凝集试验调查种畜布氏杆菌病的报告   总被引:1,自引:0,他引:1  
布氏杆菌病是一种人畜共总的传染病.主要症状表现为母盲流产、子宫炎,公言发生单九炎等病症,我市曾在工983年于某如羊场检出了羊型布氏杆菌病.为了弄清我市布氏杆菌病的疫情现状,制定出切实可行的布氏杆菌病防制措施,我们采用平板凝集和试管凝集试验对我市现有种畜场的种畜和农村种猪进行了普查,现将结果报道如下.1材料与方法1.IM#1.】.l抗原、阳性血清、阴性血清:由省畜牧兽医总站提供.l·1·2被检血清:按无菌操作采血,分离各种畜场种畜及农村种猪血清,加双抗,置0~4℃低温保存备用.互.2方法1.2.且平板凝集反应:按…  相似文献   

4.
为控制规模较小、感染率较低的山羊群布氏杆菌感染并最终净化布氏杆菌病,利用虎红平板凝集试验和试管凝集试验反复监测,及时隔离虎红平板凝集试验阳性试管凝集试验阴性羊只、扑杀并无害化处理试管凝集试验可疑或阳性羊只、停止自然交配配种。结果表明,经一定时期监测淘汰,布氏杆菌得到控制并最终得以净化。  相似文献   

5.
余荣权 《动物保健》2014,(9X):50-51
为控制规模较小、感染率较低的山羊群布氏杆菌感染并最终净化布氏杆菌病,利用虎红平板凝集试验和试管凝集试验反复监测,及时隔离虎红平板凝集试验阳性试管凝集试验阴性羊只、扑杀并无害化处理试管凝集试验可疑或阳性羊只、停止自然交配配种。结果表明,经一定时期监测淘汰,布氏杆菌得到控制并最终得以净化。  相似文献   

6.
以纯化的牛分枝杆菌重组MPT83蛋白为包被抗原,建立了检测牛分枝杆菌抗体的间接ELISA方法。确定了间接ELISA各组分的最适反应条件:抗原包被浓度为1μg/mL,酶标二抗稀释度为1:1600,血清稀释度为1:60,抗原和血清、血清和二抗均在37C反应30min,底物在37℃显色15min,D655nm阴性、阳性临界值为0.5。经阻断试验、交叉试验、重复性试验,表明该方法特异性强、重复性好。用该方法对18份结核菌素试验阳性牛血清和36份结核菌素试验阴性牛血清进行检测,结果显示,阳性血清的符合率为27.8%,阴性血清的符合率为91.7%。  相似文献   

7.
应用虎红平板和试管凝集试验对3607份来自格尔木地区牛、羊、鼠兔、旱獭、田鼠和小家鼠的血清样品进行布氏杆菌病特异性抗体的检测。结果用虎红平板检出阳性血清66份,血清阳性率为1.83%;用试管凝集试验检出阳性14份,阳性率为0.39%,说明在格尔木地区的野生动物群中存在布氏杆菌病的感染。  相似文献   

8.
为了解甘肃省兰州市城关区犬布鲁氏菌病的流行情况,从兰州市城关区动物医疗机构和兰州市流浪动物救助站共采集犬血清343份,采用光滑型虎红平板凝集试验、粗糙型虎红平板凝集试验、通用型胶体金检测法和光滑型胶体金检测法进行检测,对4种方法检测出的阳性血清进行通用型快速PCR复检;对光滑型虎红平板和粗糙型虎红平板凝集试验检测出的阳性血清再分别进行光滑型试管凝集试验和粗糙型试管凝集试验复检。结果表明,4种方法共检出34份阳性血清,其中抗体检测法虎红平板凝集试验、试管凝集试验及布鲁氏菌通用型抗体检测试纸条分别检测出阳性血清22份、18份和12份,抗体阳性率分别为6.4%、5.2%和3.5%;抗原检测法通用型快速PCR检出阳性血清25份,抗原阳性率为7.2%。22份虎红平板凝集阳性样本经光滑型试管凝集试验和粗糙型试管凝集试验复检,共检出阳性血清18份,且均为粗糙型;布鲁氏菌光滑性抗体金标快速检测卡检测均为阴性。说明兰州市城关区犬感染的主要是粗糙型布鲁氏菌,快速通用型荧光PCR的阳性检出率最高。  相似文献   

9.
为了解近几年贵州省各地区牛羊布氏杆菌病的流行情况及与温度、海拔高度等环境地理因素的相关性,采用虎红平板凝集试验和试管凝集试验相结合的方式对全省9个地州市采集4 058份(牛1 932份,羊2 126份)血清样本进行平行检测。结果:虎红平板凝集试验的血清阳性检出率,牛为14.44%,羊为3.77%;试验管凝集试验的血清阳性检出率,牛为7.66%,羊为3.17%。与后者相比,虎红平板凝集试验对牛、羊血清的敏感性均为100%,对牛血清的特异性为92.66%,符合率为93.22%;而对羊血清的特异性为99.37%,符合率为99.39%。对不同饲养方式牛羊布氏杆菌感染阳性率进行比较,总体趋势表现为牛:规模养殖场>养殖小区>农村散养户;羊:规模养殖场>农村散养户>养殖小区。通过偏相关分析环境因子与布氏杆菌感染阳性率关系为年平均温度与布氏杆菌感染抗体阳性率呈负相关,海拔高度与布氏杆菌感染抗体阳性率无相关性。  相似文献   

10.
第147部国家家禽改良计划——辅助规则 (g)做该试验应具有适合的、平滑的反应板。置备好血清抗原稀释液,以保证在将标准试管凝集试验作对照试验时稀释度不超过1:50。对于火鸡血液样本,最好使用与试管试验法相同的血清抗原稀释度(1:25)。应将血清加入抗原中,并用血清移液管顶端彻底混均抗原和血清。  相似文献   

11.
A dot Enzyme-linked Immunosorbent Assay (dot-ELISA), using whole cell Brucella abortus antigen dotted on the nitrocellulose membrane bound to a plastic strip (dipstick) was employed for the detection of Brucella antibodies in bovine sera. The results were compared with that of serum agglutination (SAT), Rose Bengal plate agglutination (RBPT) and Complement Fixation test (CFT). All the four tests gave negative reaction in 127 sera obtained from a brucellosis free herd. Testing of 549 sera from a chronically infected herd revealed 57 positive and 447 negative animals in all the four assays. Of the remaining 45 sera, 34 were positive in dot-ELISA. Six of these cases were independently detected by dot-ELISA while 28 showed positive reactions in combination with other tests. When serum samples from 158 aborted cases were subjected to dot-ELISA, 79 were found positive. Of these dot-ELISA positive cases, 71 gave positive reaction in SAT, 72 in RBPT and 78 in CFT. B. abortus biotype 3 was isolated from 34 of the 98 aborted fetuses examined.  相似文献   

12.
A total of 423 serum samples representing 94 coyotes which were wild trapped in east Texas were used to compare the serologic results from five different methods for detecting antibodies to Brucella abortus. The sera were tested for Brucella spp. antibody activity by the Card (CARD), rivanol precipitation (RIV), standard agglutination tube (SAT), cold complement fixation test (CF), and enzyme linked immunosorbent assay (ELISA) methods. Each serum sample selected for this comparison demonstrated antibody activity by one or more of the five serologic methods. When the serologic results of the five different methods were compared, 143 sera were positive according to the CF test and agreement was 67.1-70.6% with CARD, RIV and SAT. The maximum agreement for CF positive was with CARD (70.6%) and the lowest agreement fro CF negative was also with CARD (56.4%). Agreement among the serologic methods for the SAT positive ranged from 69.1% (CARD) to 72.7% (RIV). Agreement between SAT and ELISA was poor with only 38.1% agreement for SAT positive and 11.3% agreement for SAT negative. Agreement between methods for CARD positive sera was poor, with a low of 43% for both SAT and ELISA, and a high of 55.6% for RIV. Agreement between methods for 149 RIV positive sera was 83.2% for CARD, 67.8% for SAT, 64.4% for CF and only 50.3% for ELISA. Agreement between methods for ELISA positive results ranged from 49.0% for RIV to 62.7% for CARD.(ABSTRACT TRUNCATED AT 250 WORDS)  相似文献   

13.
Serial dilution and single dilution enzyme linked immunosorbent assays (ELISA) were standardised and their sensitivity and specificity were compared for serodiagnosis of Babesia equi infection. The antibody titres of 24 donkey sera of known identity were determined separately by serial dilution ELISA using three different B. equi antigens namely whole merozoite (WM), cell membrane (CM) and high speed supernatant (HSS). The ratios of the optical density (OD) of known positive and known negative sera at different serum dilutions were calculated and termed as the positive/negative (P/N) ratio. The coefficients of correlation (r) were calculated between the P/N ratios at different dilutions of sera and the log10 antibody titres of the same sera were ascertained by serial dilution ELISA. The highest value of 'r' was obtained at a serum dilution of 1:200. From log10 antibody titre of sera (y) and their P/N ratio at a dilution of 1:200 (x), regression equations (y = a + bx) were calculated separately for the three antigens. Test sera were diluted to 1:200, their OD were read in duplicate wells and were converted to the P/N ratio. Antibody titres were predicted from the P/N ratio using a regression equation separately for the three antigens. Titres obtained by both ELISAs were not significantly different from each other, thus confirming that single dilution ELISA could be successfully used to replace conventional serial dilution ELISA. The sensitivity, specificity and predictive value of single dilution ELISA was validated statistically using 42 B. equi disease-positive sera and 106 B. equi disease-negative sera. The WM antigen was found to be the most sensitive with a higher predictive value for negative test sera as compared to the CM or HSS antigens. Sera positive for other equine infections including Babesia caballi showed no cross-reaction with the three B. equi antigens in ELISA, thus the test was immunologically specific. Antibody titres of 109 unknown field donkey/horse sera obtained by serial and single dilution ELISA using the WM antigen did not show any significant difference. Since the single dilution ELISA was found to be more economical, convenient, sensitive, specific than the serial dilution ELISA and has a high predictive value, it is suitable for use in sero-epidemiological studies on B. equi infections in the field.  相似文献   

14.
A recombinant leptospiral lipoprotein, LigB, was evaluated for use in the diagnosis of bovine leptospirosis by enzyme-linked immunosorbent assay (rLigB IgG ELISA). The standard reference test (Microscopic agglutination test, MAT) of 200 serum samples from cattle suspected of leptospirosis showed that 95 (47.5%) samples had positive agglutination titres, which ranged from 100 to 1600. In rLigB IgG ELISA, 49% of the samples were positive. Sensitivity of IgG ELISA for 95 bovine sera, which had MAT titres of greater than or equal to 100, were 100%. ELISA showed a specificity of 97.1% with 105 bovine sera, which were negative at a 1:50 dilution in MAT for Leptospira interrogans serovars. The results of ELISA and MAT correspond very good. When analytical specificity of IgG ELISA was evaluated using bovine serum samples from animals showing the serum antibodies to other pathogens, no cross-reaction was observed. Thus the recombinant LigB IgG ELISA can be used instead of the MAT as an aid to the diagnosis of bovine leptospirosis.  相似文献   

15.
An enzyme-linked immunosorbent assay (ELISA) was evaluated and compared in parallel with the standard complement fixation test (CFT) for the diagnosis of bovine subclinical paratuberculosis. Bovine sera preabsorbed with the mixture of Mycobacterium phlei and kaolin suspension were assayed for antibody activities to the crude protoplasmic antigen of Mycobacterium paratuberculosis in the ELISA. ELISA antibody titer was expressed as ELISA antibody index (EAI) value: EAI = (At-An)/(Ap-An), where At, Ap and An are the absorbance values of a 1:200 dilution of unknown test sera, a 1:400 dilution of positive control serum, and a 1:200 dilution of negative control serum. An EAI of 0.6 or greater was established as a reasonable cutoff point for a positive antibody titer by ELISA. Of the 156 sera from cattle with subclinical M. paratuberculosis-infection, 106 (67.9%) were positive by ELISA and 41 (26.3%) by CFT. Of the 3,880 sera from cattle in the herds which had no history or evidence of paratuberculosis, 3,875 (99.9%) were negative by ELISA, and 3,787 (97.6%) by CFT. Positive ELISA titers were detectable 1 to 5 months earlier than positive CFT titers in experimentally infected cattle, and 7 to 10 months earlier in naturally infected cattle. These results indicate that the ELISA should replace the CFT as the routine test of choice for the diagnosis of bovine paratuberculosis.  相似文献   

16.
副猪嗜血杆菌间接ELISA抗体检测方法的建立   总被引:15,自引:0,他引:15  
采用福尔马林灭活的副猪嗜血杆菌全菌体作为包被抗原,建立了检测副猪嗜血杆菌抗体的间接ELISA方法。经试验确定副猪嗜血杆菌全菌体的包被浓度为2·24×107CFU/孔、检测血清为1∶200稀释,同时确立了间接ELISA的最佳反应条件。该方法有很高的特异性和重复性,14个发病猪场100份血清检测结果Hps抗体阳性检出率为94%,明显高于细菌分离鉴定检测结果。  相似文献   

17.
Outer sheath antigen was prepared from Leptospira interrogans serovars pomona, sejroe and hardjo by treating the organisms with 1.0M NaC1 followed by 0.04% sodium dodecyl sulfate (SDS). Sodium dodecyl sulfate was removed from the SDS-protein complexes by the extraction of dodecyl sulfate anions as ion pairs with triethylammonium cations into an organic solvent. The outer sheath antigen was recovered from the organic solvent as a precipitate and used as the source of leptospiral enzyme-linked immunosorbent assay (ELISA) antigen. Utilizing this antigen, ELISA was adapted to detect bovine serum antibody to L. interrogans serovars pomona, sejroe and hardjo. The specificity of this assay in 344 bovine sera, which were negative in the microscopic agglutination test (MAT) for seven serovars, was 99.4%. In sera from 37 and 87 cattle which revealed MAT titers greater than or equal to 1:50 for L. interrogans serovars pomona and sejroe, the relative sensitivity of the test was 100%. The ELISA also showed a considerable degree of low level cross-reactivity with other serovars. Sixty-six (75.9%) out of 87 bovine sera which were MAT-positive (MAT titer of greater than or equal to 1:50) with serovars sejroe and hardjo only were ELISA positive with heterologous pomona antigen; 16 (43.2%) and six 16.2%) out of 37 bovine sera which were MAT positive MAT titer of greater than or equal to 1:50) with serovar pomona only were ELISA positive with heterologous sejroe and hardjo ELISA antigen respectively.(ABSTRACT TRUNCATED AT 250 WORDS)  相似文献   

18.
The serum agglutination test (SAT), the Rose Bengal plate test (RBPT) and the milk ring test (MRT) were used in the diagnosis of caprine brucellosis. There was a close correlation between the SAT and RBPT when both tests were negative but the RBPT failed to detect 79.82 per cent of sera in excess of 50 iu. Also, owing to the relatively poor milking potential of the Nigerian goat and the false positive results with the MRT, it is concluded that the SAT offers a better serological diagnostic tool for caprine brucellosis in this locality.  相似文献   

19.
In this study, Brucella antibodies in bovine sera and milk were detected using the dot-immunobinding assay (DIA), the serum agglutination test (SAT), the Rose Bengal plate test (RBPT) and the milk ring test (MRT). For this purpose, a total of 116 paired blood and milk samples collected at the same time from 56 aborted and from 60 healthy dairy cows was examined. In DIA, a nitrocellulose membrane (NCM) was used as the solid phase. Antigen adsorbed on the NCM was extracted from Brucella abortus S99 by heat treatment. The results obtained by DIA were compared with those of SAT, RBPT and MRT. Of the 116 paired blood and milk samples, 24 were positive and 72 were negative by all tests used. Serum samples of six aborted cows were positive by DIA, SAT and RBPT but the milk samples were negative by DIA and MRT. Serum and milk samples of four aborted cows gave positive reaction only by DIA tests. The remaining six aborted cows were negative only by MRT and two of them were negative by both RBPT and MRT. Four sera of healthy cows were found to be positive only by SAT.  相似文献   

20.
为满足口岸对南非Ⅱ型口蹄疫检测、监测的需求,在对南非Ⅱ型口蹄疫抗原表位分析及相关基因合成的基础上,进行融合蛋白pGEX-VP1~VP3的体外表达,采用柱上酶切方法制备VP1~VP3蛋白。将纯化VP1~VP3抗原包被96微孔板,分别优化抗原包被浓度、血清稀释度、酶标二抗稀释度,确定阴性临界值。经反复优化,建立的南非Ⅱ型间接ELISA检测方法的最佳抗原包被浓度为6.4ug/ml,血清最佳稀释度为1:200,酶标二抗最佳稀释度为1:4000,阴性临界值(OD450)为0.562。检测样品判定标准为:OD450>0.622为阳性,OD450<0.502为阴性,0.502≤OD450≤0.622为可疑。特异性试验结果表明,所建立方法与其它血清型口蹄疫间无交叉反应。南非Ⅱ型口蹄疫间接ELISA检测方法的建立为我国口岸口蹄疫检疫提供了新的查验方法。  相似文献   

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