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1.
应用琼脂扩散试验(AGP)法对具有典型临床症状并经病理学诊断确诊的鸡血清50份和法氏囊材料50份进行了标准抗原检测被检血清;标准阳性血清检测被检疫抗原的研究。标准抗原检测被检血清阳性40份,阳性率为80%(40/50);标准阳性血清检测被检抗原阳性36份,阳性率为72%(36/50)。经x^2检验(P>0.05)差异不显著。结果表明两者均适应于IBD的诊断。  相似文献   

2.
采用本研究室制备的3批传染性支气管炎病毒(IBV,M41株)HI试验抗原,分别对100份和80份不同鸡血清进行IBV HI抗体检测,同时用IBV ELISA抗体检测试剂盒进行检测,比较两种不同方法检测的特异性和敏感性。结果显示,特异性试验中80份SPF鸡血清,自制抗原检测均为阴性,IBV ELISA检测79份为阴性,10份其他鸡病血清,两种方法检测9份均为阴性,10份IBV阳性血清两种方法检测均为阳性;敏感性试验中,74份已知IB疫苗免疫或IBV M41株感染鸡血清IBV HI检测72份为阳性,阳性检出率为97.3%(72/74),IBV ELISA检测74份均为阳性,阳性检出率为100%(74/74),SPF鸡血清及其他鸡病血清,两种方法检测均为阴性,两种检测方法总符合率为97.5%,差异不显著(P0.05)。试验结果证明,本研究室自制抗原具有良好的特异性,特异性为100%,抗原同时具有良好的敏感性,但IBV ELISA方法的敏感性要高于IBV HI方法。  相似文献   

3.
在应用PPA—ELISA间接法检测兔病毒性出血症血清抗体的基础上,将标准阳性血清用肝脏抗原进行吸收,以标准阳性血清OD值的变化来判定有无肝脏抗原。本试验共检测血清171份,肝脏57份。试验结果表明,本方法简便、特异性强、灵敏度高、结果便于判定。  相似文献   

4.
为了探讨用抗独特型抗体替代病毒或病毒亚单位检测H9亚型禽流感血清抗体的可行性,本试验分别用全病毒抗原和抗独特型抗体(Ab2)作检测原,通过琼脂免疫扩散试验(AGP)和间接酶联免疫吸附试验(ELISA)检测了鸡的血清样品。在AGP试验中,分别用Ab2和禽流感病毒(AIV)对10份血清样品检测的符合率为70%。在间接ELISA试验中,Ab2和AIV抗原对10份血清样品检测的符合率为90%。用Ab2间接ELISA试验检测出的血清阳性率(8/10)高于血凝抑制试验(6/10)和AGP试验(5/10)。结果表明,在一定的情况下Ab2可以用来代替具有污染环境风险的病毒抗原来检测抗病毒抗体。  相似文献   

5.
用琼脂凝胶双扩散试验发现了囊虫病猪的循环抗原,并用反向间接血凝技术作了检测。103份囊虫病猪血清琼扩试验对循环抗原的检出率为22.33%(23/103);反向间接血凝试验检出率为32.03%(33/103),其余70份呈阴性反应的血清经免疫复合物解离后,又有25份呈阳性反应,总检出率为56.31%(58/103)。  相似文献   

6.
建立了检测包虫病循环抗原的夹心酶联免疫吸附试验。对130份阳性血清(猪65,羊65)作了检测,总阳性率为79.2%,抗原最小检出量为50μg/L。  相似文献   

7.
用培养的布氏杆菌菌体,通过超声波裂解、反复离心制备出布氏杆菌细胞壁抗原。将细胞壁抗原作1:128稀释,用作牛种布氏杆菌酶联免疫吸附试验(ELISA)的抗原;将布氏杆菌细胞壁抗原作1:32稀释,用作平板凝集试验抗原;把细胞壁抗原作1:16稀释用作试管凝集试验抗原,分别建立了牛布氏杆菌ELISA试验、平板凝集试验和试管凝集试验。用这3种方法,检测已知200份平板凝集试验阴性血清,5份平板凝集试验阳性血清。结果5份阳性血清在平板凝集试验、试管凝集试验和ELISA试验中均为阳性;200份阴性血清在平板凝集试验和试管凝集试验中均为阴性,在ELISA试验中有1份为阳性?试验证明,细胞壁抗原,既能用于传统的平板凝集试验和试管凝集试验,又能用于ELISA试验。  相似文献   

8.
以羊口疮痂皮毒抗原接种家兔制备高免血清,经琼扩试验检测血清价为1∶16;通过抗原与兔抗羊口疮高免血清的方阵分析,确定兔抗羊口疮高免血清诊断抗体的最佳稀释度为1∶4;该诊断抗体对5份不同地区的羊口疮疑似病料都呈现特异性反应,不与山羊痘疫苗毒、口蹄疫疫苗毒和羊正常皮肤抗原发生交叉反应。琼脂扩散试验检测羊口疮病毒是一种简便、易于判断、实用性强的诊断方法。  相似文献   

9.
研究旨在调查南疆某规模化猪场免疫猪群的猪瘟抗原与抗体水平,为制定适应当地生产条件的免疫程序提供参考。试验运用酶联免疫吸附试验(ELISA)对该场免疫猪群的480份血清样品进行抗体检测与分析,并采用反转录-聚合酶链式反应(RT-PCR)对采集的24份猪瘟疑似病料进行抗原检测。结果显示,对随机采集的480份血清样品进行猪瘟抗体检测,其中449份为阳性样品,阳性率为93.5%;不同类别群体中以母猪的抗体阳性率最高,为97.2%;2~4胎母猪与5~6胎母猪的抗体阳性率为97.8%;22~28 d的阳性率最低,为76.9%;36~50 d的仔猪抗体阳性率最高,为92.3%。对采集的24份猪瘟疑似病料进行抗原检测,显示均为阴性。研究表明,该规模化猪场的猪瘟免疫情况良好,未出现感染情况。  相似文献   

10.
应用对流免疫电流技术(CIET)和琼脂扩散试验(AGP)对具有典型临床症状传染性法氏囊病鸡血清50份和法氏囊材料50份进行标准抗原检测被检血清,标准阳性因清检测被检抗原。AGP法标准抗原检测被检血清阳性40份,阳性率80%(40/50),标准阳性血清检测被检抗原,阳性36份,阳性率72%(36/50);CIET法标准阳性血清检测被检抗原,阳性47份(47/50),阳性率为94%,标准抗原检测被检血清,阳性48份,阳性率为96%(48/50)。经x^2检验均P>0.05,差异不显著。CIET法与AGP法,标准抗原检测被检血清,AGP法阳性率80%(40/50),CIET法阳性率96%(48/50);标准阳性血清检测被检抗原,AGP法阳性率72%(36/50),CIET法阳性率94%(47/50)。经x^2检验均P<0.05,差异显著。结果表明,CIET法比AGP法敏感、快速、且检出率高。  相似文献   

11.
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   

12.
牛副结核ELISA诊断方法研究   总被引:5,自引:0,他引:5  
本文建立了牛副结核病ELISA诊断方法。采用亲和层析抗原。被检血清以高压粉碎草分枝杆菌吸收原进行吸收。该方法的敏感性76%,特异性97%,通过对2483头奶牛进行检测,检出率为6.1%,并与常规的补反和变态反应进行了比较。作者认为:牛副结核ELISA可以替代补反,做为检测牛副结核的一种有力手段。  相似文献   

13.
An enzyme-linked immunosorbent assay (ELISA) was adapted to detect Mycoplasma californicum-specific antibodies in bovine serum. Cross-reactive antibody was found in the M californicum-positive reference serum when assayed against each of 7 solid-phase antigens of heterologous mycoplasma species. Cross-reactivity was further demonstrated by inhibition of ELISA reactivity to M californicum solid-phase antigen by incubation of sera with antigen suspensions of each heterologous species. Incubation of test sera with a cross-reacting antigen mixture containing equal proportions of the 7 cross-reactive mycoplasmas was used to minimize cross-reactivity in the M californicum-specific ELISA. Specificity of antibody reactivity to M californicum, as measured by ELISA, was determined by enzyme-linked immunosorbance inhibition, in which sera were incubated with M californicum antigen suspensions before determining ELISA reactivity to M californicum solid-phase antigen. Seropositive and suspect sera (n = 55) were obtained from 3 dairies that had bacteriologically verified epizootics of M californicum mastitis. The percentage of inhibition demonstrated in enzyme-linked immunosorbance inhibition was determined for each serum. Inhibition percentages below the 15th percentile (61% inhibition) of this distribution were classified as nonspecific.  相似文献   

14.
A reliable method was developed to produce a viral antigen preparation from porcine reproductive and respiratory syndrome virus (PRRSV) infected MARC-145 cells by solubilizing the virus with Triton X-100. This method eliminated problems previously encountered with high background reactions associated with PRRSV antigen or cell control antigen. With this new antigen, an indirect enzyme-linked immunosorbent assay (ELISA) was adapted to detect swine serum anti-body against PRRSV. In the ELISA, non-specific reactions associated with test serum samples have been eliminated by utilizing an effective blocking serum diluent. The ELISA is more sensitive than an indirect immunofluorescent assay (IFA), particularly with late-infection sera, while maintaining a high diagnostic specificity. In a comparison of IFA and ELISA using sera collected from 250 pigs of various ages from 5 herds that had PRRS histories, IFA revealed 178 positive samples and 72 negative samples. All of the IFA-positive sera were proven to be ELISA reactors. However, nearly one-half (34/72) of the IFA-negative samples were also ELISA reactors. The diagnostic specificity and sensitivity of the ELISA were 100% and 96.6% with 257 serum samples collected from known healthy PRRS-negative swine herds and 57 sera collected from infected swine at 6 to 56 days after infection, respectively. The ELISA is technically superior to IFA, time-efficient and cost-effective, and suitable for testing of a large number of samples over a short period of time.  相似文献   

15.
A liquid-phase enzyme-linked immunosorbent assay (ELISA) was developed for the detection of vesicular stomatitis virus (VSV) types New Jersey (VSV-NJ) and Indiana subtype Indiana I (VSV-IND1) antibodies in the sera of naturally and experimentally infected cattle, horses and swine. Four different VSV preparations were compared for use as antigen in the ELISA: virus used in neutralization tests, complement-fixation antigen, immunodiffusion ager gel antigen and viral glycoprotein. Comparative antibody titration results of virus neutralization (VN) and ELISA showed no statistically significant difference between serum titers obtained with the four antigens to VSV-NJ (P=0.21) and VSV-IND1 (P=0.14). The viral glycoprotein antigen was incorporated in the ELISA system because it is non-infectious and induces neutralizing antibodies. The reliability and variability of the ELISA was determined by testing 516 bovine, equine and swine sera which originated from areas free of vesicular stomatitis, and by testing 186 sera from areas where outbreaks occur. ELISA and VN results were correlated (P < 0.001 for both serotypes), and no statiscafically difference was found between replications of the tests. The ELISA allows the testing of a larger number of sera in a shorter time than is possible with the VN test and it can be used in diagnostic laboratories in VSV-free areas for the support of epidemiological surveillance programs.  相似文献   

16.
An enzyme-linked immunosorbent assay (ELISA) for detecting serum antibodies to the porcine epidemic diarrhea coronavirus (PEDV) was established by using cell culture-grown PEDV as antigen for coating. Ultracentrifugation through 20 and 45% (w/w) sucrose cushions proved to be the best antigen purification method. Examination of 1024 swine sera showed a high specificity and a greater sensitivity of the ELISA, when compared with indirect immunofluorescence. Reference sera with high antibody titers to PEDV originated from two pigs experimentally infected with PEDV. Three different antigen purification methods and the advantages of the ELISA compared with an immunofluorescence test are discussed.  相似文献   

17.
A reliable bovine viral diarrhea (BVD) viral antigen was prepared from BVD virus grown on Madin Darby bovine kidney (MDBK) cells by solubilizing the virus with detergent MEGA-10 (decanoyl-N-methylglucamide) followed by removal of hydrophobic proteins with Triton X-100 treatment. By these treatments, problems of high background associated with BVD viral antigen in the enzyme-linked immunosorbent assay (ELISA) were eliminated. With this new antigen, an ELISA was adapted to detect bovine serum antibody against BVD virus. The diagnostic specificity of the assay in 403 bovine sera collected from a BVD virus-free herd was 100%; in 296 bovine sera with serum neutralizing antibody titers of greater than or equal to 1:2, 289 sera were ELISA positive (relative sensitivity of 97.6%), two sera gave false negative reactions (0.7%) and five sera gave suspicious reactions (1.7%). These interpretations were based on positive/negative (P/N) ratio readings, i.e. a P/N ratio of less than 1.50, 1.50-1.99 and greater than or equal to 2.00 were interpreted as negative, suspicious and positive reactions, respectively. The ELISA results gave excellent agreement with serum neutralization in detecting both seropositive and seronegative animals (Kappa = 0.994). The ELISA assay was considered to be technically superior to the serum neutralization test for the routine detection of BVD viral antibody in bovine sera.  相似文献   

18.
A simple and inexpensive method of antigen preparation by ultrafiltration was investigated using the V4 strain of Newcastle disease virus. The antigen designated XM300 was used in an indirect enzyme-linked immunosorbent assay (ELISA) for the detection of antibodies to Newcastle disease virus in chicken serum. The assay was evaluated using both experimental and field sera, as well as reference control reactor and non-reactor sera. Antigen prepared by the ultrafiltration method was compared with antigen prepared by ultracentrifugation and the ultrafiltration antigen was found to react specifically with Newcastle disease virus antiserum in this ELISA system. This antigen preparation technique is also suitable for use in developing countries. The ELISA provides an excellent method for measuring antibodies in the early stages of infection in serum samples from experimentally infected chickens. More than 14.58 % of the total serum samples which failed to be recognized as reactors by the conventional haemagglutination inhibition test were detected in the ELISA.  相似文献   

19.
An enzyme-linked immunosorbent assay (ELISA) for the survey or titration of bovine sera for the presence of IgG antibodies against infectious bovine rhinotracheitis (IBR) virus was developed. The optimal conditions of serum dilution, antigen concentration, conjugate dilution, substrate concentrations, and reaction time were established using the signal/ noise (S/N) ratio as the determining criterion. Equilibrium density gradient purified IBR virus was used as antigen at an optimal concentration of 0.60 μg/cuvette. The use of purified antigen allowed the testing of sera at a 1 : 10 dilution without nonspecific reaction.The conditions of conjugate dilution, substrate concentration and reaction time were shown to have significant effects on the ELISA test. Results from 35 sera showed this optimized ELISA procedure to be as much as 1000-fold more sensitive than the serum neutralization plaque reduction assay. Numerous sera showing no neutralizing titer to IBR virus were found to be positive when examined by this ELISA method.  相似文献   

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