酶联免疫吸附试验（ELISA）与平板凝集反应（RBPT）检测鹿布氏杆菌病…

在１９９５－１９９７年间本文作者进行了ＥＬＩＳＡ法与平板检测鹿丰氏杆菌病的研究，研究结果表明：ＥＬＩＳＡ法比平板法敏感。通过现场３００头分鹿血清的检测结果表明，前者比后者的检出率高５．３４％。前者的阳性血清中完全一者的阳性血清，同时在后者的可疑和阴性血清中，也可检出ＥＳＩＡＡ阳性。     

沈阳地区鹿结核病和布鲁菌病的血清学调查

为了解结核病和布鲁菌病在沈阳地区鹿群中的流行传播状况,从沈阳地区随机采集鹿血清样本1 055份,分别用平板凝集法检测鹿布鲁菌病和间接ELISA法检测鹿结核病的血清阳性率.经检测,沈阳地区的鹿群中结核病的血清阳性率为99,24％,鹿布鲁菌病的血清阳性率为20.19％.本次调查结果为沈阳地区鹿结核病和布鲁菌病的防控提供了流...     

细胞壁抗原用于布氏杆菌病诊断试验的研究

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。     

牛布鲁氏菌病不同检测方法比较与试剂质控

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。     

用PPA-ELISA间接法检测兔病毒性出血症(RHDV)肝脏抗原及血清抗体的研究

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。     

酶联免疫吸附试验诊断黄牛布氏菌病(摘要)

<正> 用酶联免疫吸附试验(ELISA),对139例黄牛血清标本,进行了布鲁氏菌病的抗体检测,阳性标准是1:320。同时与常规的试管凝集试验(SAT)、半胱氨酸凝集试验(CAT)、平板凝集试验(PAT)、虎红平板凝集试验(RBPT)四种试验方法进行了对比和分析。     

J亚群禽白血病病毒(ALV-J)ELISA检测方法的建立

利用抗J亚群禽白血病病毒（ALV—J）囊膜蛋白特异性单克隆抗体JE9，建立了检测ALV—J env抗原抗体免疫复合物的ELISA方法。应用该方法检测SPF鸡血清、鸡抗禽流感H9亚型阳性血清、鸡沙门氏菌阳性血清、鸡腺病毒阳性血清、鸡新城疫阳性血清．结果均为阴性，无交叉反应；抗ALV—J阳性血清与ALV—J特异性单克隆抗体JE9能相互阻断；ALV—J阳性血清经酸处理后，ELISA检测的D490差值明显下降。对部分攻毒鸡血清样本及田间ALV—J阳性血清样本进行电镜观察，可见ALV—J样病毒粒子及ALV—J样免疫复合物。经与间接免疫荧光（IFA）检测ALV—J env抗体结果比较表明，建立的ELISA方法与IFA方法两者具有较好的群体符合率，群体符合率为8／9。这些结果表明，本研究建立的ELISA方法在ALV—J的诊断中具有很好的应用前景。     

猪乙型脑炎间接ELISA与血凝抑制试验方法的比较

用血凝抑制试验和间接ELISA两种方法检测乙脑阳性血清均呈阳性反应,检测乙脑阴性血清均呈阴性反应。用间接ELISA对来自9个猪场74份送检血清进行了猪乙脑病毒（Japanese encephalitis virus,JEV）抗体检测,并与血凝抑制试验（HI）进行对比,两种方法检测总符合率为85.1%(63/74),经统计分析,检测结果差异不显著(P＞0.05)。对来自无猪乙脑病猪场24头健康猪的血清进行检测,两种方法结果均为阴性,符合率为100%。对10份阳性血清进行检测,ELISA检测效价较HI效价高,表明ELISA比HI敏感。结果表明,ELISA与HI检测结果相符,前者更适用于猪血清中乙脑IgG抗体的大规模检测。     

F107(F18ab)菌毛单抗的制备、鉴定与初步应用

将大肠杆菌107／86和水肿病菌株在TSB中培养，鉴定其表达F107菌毛，前者作为免疫原免疫BABL／C小鼠，后者作为检测抗原包装酶标板。细胞融合后经ELISA和玻板凝集检测，得到一株阳性杂交瘤细胞株，命名为F7－1。该细胞株制备的腹水ELISA效价为：2^16，试管凝集价为：1：5120，在免疫印迹中可与提取的F107菌毛特异性条带发生反应。利用此单抗检测表明，91％的水肿病菌株表达F107菌毛。     

全血与血清平板凝集试验检测鸡白痢的对比

对北京油鸡两个品系216只母鸡,在17~49周期间每隔8周同时用全血和血清平板凝集试验进行鸡白痢检测对比分析。结果发现,血清检出阳性率高于全血检出阳性率,且血清检出阳性个体包含全血检出阳性个体;同时,检测结果也表明,33周龄鸡白痢阳性率达到最高,此后阳性率下降。因此,在鸡白痢净化过程中采用血清平板凝集法比全血平板凝集法检测更为敏感,在25~33周对北京油鸡群体增加一次鸡白痢净化更有助于加快净化进程。本研究可为其他地方品种种鸡鸡白痢检测工作提供参考。     

奶牛布氏杆菌病的血清学诊断

为了解虎红平板凝集试验(RBPT)、试管凝集试验(SAT)和酶联免疫吸附试验(ELISA)三种血清学方法在奶牛布氏杆菌病诊断方面的实际应用情况,本试验采集了10份疑似奶牛布氏杆菌病的血清样本,同时采用上述三种血清学诊断方法进行了对比研究。结果显示,RBPT、SAT和ELISA的阳性检出率分别为70%、80%和90%,以ELISA的阳性检出率为最高。因此,具备特异、灵敏、快速等优点的ELISA血清学诊断方法更适合奶牛布氏杆菌病的抗体检测及其研究。     

鉴别布鲁菌病自然感染动物与疫苗免疫动物胶体金检测试纸条的研制

用金黄色葡萄球菌A蛋白（SPA）标记胶体金技术制作金标垫,以亲和层析法纯化的BP26蛋白作为检测抗原（T线）,研制胶体金快速检测试纸条。用建立的试纸条检测小鼠布鲁菌感染血清,可成功鉴别牛布鲁菌S19感染和BP26缺失疫苗株YZ-2免疫的小鼠。试纸条检测与布鲁菌属同源性较近的几株细菌的阳性血清,结果无交叉反应;试纸条敏感性高于虎红平板凝集试验检测方法,近似于ELISA方法;准确性同于ELISA法,且更为简便快捷。该试纸条具有鉴别布鲁菌病自然感染和疫苗免疫的应用前景。     

虎红平板凝集试验与试管凝集试验检测奶牛布鲁菌病的比较

布鲁菌病主要采用凝集试验来检测,本试验用虎红平板凝集试验和试管凝集试验对江苏海安、江都、无锡和兴化地区的660头奶牛的血清样品进行平行检测,虎红平板凝集试验的阳性检出率为9.7%,试验管凝集试验的阳性检出率为5.5%;与后者相比,虎红平板凝集试验的敏感性为100%,特异性为95.3%,符合率为95.6%。这些数据表明,江苏地区某些奶牛场存在布鲁菌感染,虎红平板凝集试验可以用于布鲁菌病初步检测。     

补体结合酶联免疫吸附试验方法的建立

为改进免疫学诊断技术的准确性,研究了一种基于补体结合的免疫学检测新技术———补体结合酶联免疫吸附试验（CF-ELISA）。CF-ELISA技术采用酶标记抗菊糖纯化豚鼠补体C3抗体及其酶显色系统作为补体参与反应的指示系统,用ELISA方法进行补体结合试验。经对布氏菌病抗体检测的初步试验结果显示,CF-ELISA技术可检测到0.01 IU的布氏菌病抗体,灵敏度与间接酶联免疫吸附试验（iELISA）相当,是虎红平板凝集试验（RBPT）试管凝集试验（SAT）的5 000倍、补体结合试验（CFT）的10 000倍。对349份确诊布氏菌病感染群牛、羊血清的检测结果显示,CF-ELISAi、ELISA、CFT、SAT、RBPT的阳性率分别为35.82%3、6.39%、31.81%、30.09%、36.1%,CF-ELISA与iELISA、CFT、SAT、RBPT的阳性符合率分别为：98.4%、88.8%、80.0%、90.6%。CF-ELISAi、ELISA、CFT、SAT、RBPT对490份布氏菌病阴性群牛、羊血清的阴性率分别为100%、99.6%、100%、99.4%、99.8%,CF-ELISA与iELISA、CFT、SAT、RBPT的阴性符合率分别为：99.6%1、00%、99.4%、99.8%。研究表明,CF-ELISA是具有高特异性和高敏感性的布氏菌病免疫学检测技术。     

青海省互助县某养殖场部分母羊流产病因的检测

通过虎红平板凝集试验（RBPT）、弓形虫和衣原体间接血凝试验（IHA）,对来自青海省互助县某羊场的19份母羊血清进行了布鲁氏菌、弓形虫和衣原体的血清抗体检测.结果检出衣原体阳性血清1份,阳性率为5.3％;未检出布鲁氏菌和弓形虫的阳性血清.流产胎儿瘤胃液,经PCR检测,为衣原体感染.     

祁连县绵羊群中流产类疾病的血清学检测

应用虎红平板凝集试验(Rose-Bengal Plate Agglutination Test,RBPT)、衣原体、弓形虫、绵羊支原体间接血凝试验(Indirect Hemagglutination Assay,IHA),对采自青海省祁连县两个村绵羊的199份血清进行了布氏杆菌、衣原体、弓形虫的血清抗体检测。结果:在199份被检血清中,未检出布氏杆菌和衣原体的血清阳性;检出弓形虫阳性血清9份,阳性率为4.5%;羊传染性胸膜肺炎阳性血清13份,阳性率为6.5%。     

青海省海南州引进种牛布鲁氏菌、衣原体和弓形虫等病血清学检测

应用布鲁氏菌病虎红平板凝集试验、衣原体和弓形虫IHA试验,对青海省海南州某核心种牛群引进的40只种公牛,进行了布鲁氏菌、衣原体和弓形虫等病的特异性血清抗体检测。结果:检出布氏杆菌阳性血清1份,阳性率2.50%:检出衣原体阳性血清6份,阳性率为15.00%;弓形虫阳性血清2份,阳性率为5.00%。结果表明引进的种牛中存在不同程度的布鲁氏菌病、衣原体及弓形虫病。而该三种病均可通过垂直传播。所以,对种牛引进因加强检疫及管理等措施。防止种牛感染造成的疫病传播。     

The use of homologous antigen in the serological diagnosis of brucellosis caused by Brucella melitensis

In the European Union the serological diagnosis of brucellosis caused by Brucella melitensis is performed using the heterologous antigen of B. abortus S99. The possible higher sensitivity or ability of an early detection of antibodies by a homologous antigen may prove very useful in the final phases of an eradication programme. Results obtained in sheep experimentally infected by B. melitensis biovar 3 were compared using B. abortus S99, B. melitensis M1, M2 and M3 antigens in the Rose Bengal plate test (RBPT), the complement fixation test (CFT) and an enzyme-linked immunosorbent assay (ELISA) test. Forty-six sheep from an officially brucellosis-free flock were experimentally infected intraconjunctivally with B. melitensis biovar 3. Prior to infection, all animals were tested first against Brucella antibodies, weekly for 2 months post-infection (PI) and then monthly for a further 7 months. All sera were tested against the antigens listed above using RBPT, CFT and ELISA. Using a Bayesian approach, test sensitivities were estimated and compared. Their ability for the early detection of antibodies was evaluated through a regression model based on a logit response model, using the number of days PI as the independent variable and the logit of the fraction of positive animals as the dependent variable. No significant differences were detected among the various antigens used, either in terms of sensitivity or in terms of antibody kinetics; however, the CFT was significantly less sensitive than the RBPT and ELISA and it also showed a lower rate of increase of percentage positive animals (beta-coefficient of regression analysis).     

青海省三角城种羊场部分种羊中流产类疾病的血清学检测

应用布鲁氏菌病RBPT(Rose-Bengal Plate Agglutination Test)、衣原体和弓形虫IHA(Indirect Hemagglutination Assay),对来自青海省三角城种羊场的59份试情公羊的血清、60份后备母羊的血清进行特异性血清抗体的检测。结果:在119份被检血清中,检出衣原体阳性血清3份,阳性率为2.52%,其中在59份试情公羊血清中检出3份,阳性率5.08%;在后备母羊的血清未检出衣原体阳性血清;所有的血清中均未检测到布氏杆菌和弓形虫阳性血清。     

Control measures in officially acknowledged brucellosis-free and leukosis unsuspected dairy herds on the basis of bulk milk samples in combination with ELISA tests

1. EC- and National Regulations. Since 1988 the EC-regulations accept in addition to the on Agar Gel Immunodiffusion test (AGIDT) based blood serum testing of cattle herds that are filed as "free from Enzootic Bovine Leucosis" the use of ELISA for this purpose. The regular testings in dairy cattle herds can be done alternatively with single or pooled milk samples, in other herds with pooled blood sera using ELISA. General condition is only a minimal sensitivity of the test to detect the European EBL Antibody Standard ("E4") in a dilution of 1:10 in negative serum or 1:250 in negative milk. Adequate national regulations are in preparation. The present limitation of pool sizes, blood maximum 50 animals without preparation steps 20, and milk after concentration treatment 50 cows is neutralized by proceedings in development of higher sensitive ELISA tests. This limitation should be canceled. Herd bulk milk samples without size limitations are accepted to be tested with "Milk Ring Test" by EC for the regular testings in filed "Brucellosis Free Dairy Cattle Herds". The alternative use of more sensitive (and more specific) ELISA tests for this purpose including the technical conditions is in a final discussion. 2. Scientific-Technical Base for Using the Chances of the Proceeding in the EC-Regulations. The realisation of the EC accepted or final discussed ELISA based bulk milk testing to control filed "EBL- and/or Brucellosis Free Herds" depends on some basic conditions like sensitivity, specificity, and variability of the ELISA systems. Field trials of more than 20,000 bulk milk samples in case of Brucellosis and more than 2,000 in case of EBL show the feasibilities and the limits of the ELISA systems in defining the status of the herds. The Brucellosis respectively the EBL situations of the dairy cattle herds tested in this trail were well known by history and by investigation of single animal blood samples using conventional tests. Special test run variations of pretested assays demonstrated the possibilities to define the EBL status of dairy cattle herds up to 50 lactating cows without preparation of the bulk milk sample and up 100 after concentration of the antibodies by the rennet-ammonium sulfate method. The concentration limit for detection of Brucellosis antibodies is 100 lactating cows. The bulk milk of smaller herds can be tested without concentration. On principle the evaluation of the test values bases on defined relations to a "weak positive" reference.(ABSTRACT TRUNCATED AT 400 WORDS)