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类鼻疽快速诊断的研究 总被引：4，自引：0，他引：4
对实验感染类鼻疽杆菌的豚鼠，以间接血凝试验检测证实，类鼻疽杆菌外毒素抗体于感染后４ｄ即可测得，１６～３２ｄ达高峰；而类鼻疽杆菌脂多糖抗体出现较慢，但维持时间长。用测定外毒素抗体的间接血凝试验对７例细菌分离阳性患者进行测试，以血清滴度≥１∶８０为临界值，其敏感性达１００％；对７０８名健康者测试，其特异性达９９％。在疫区，从感染性疾病中检出类鼻疽患者，测定外毒素抗体的间接血凝试验有重要的实用价值 相似文献
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应用微量中和试验(MT)、间接血凝试验(IHA)和间接酶联免疫吸附试验(间接ELISA)对三种不同途径即滴鼻、口服和肌肉注射人工感染猪流行性腹泻病毒的12头仔猪的血清抗体进行了检测。感染猪在第1～2周时即可测出抗体,3～5周后抗体滴度达高峰,抗体持续时间最短为18周,最长可达22周以上。其中以肌注和滴鼻组抗体产生较早,而口服组稍晚。肌注组抗体上升较快,抗体持续时间较短,而滴鼻和口服组抗体上升稍慢,抗体持续时间较长。 相似文献
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根据10头人工感染弓形虫病猪的间接血凝试验（IHAT）和酶联免疫吸附试验（ELISA）血清抗体检测结果表明．所有被检猪感染后第14天血清抗体达到阳性滴度．第21天抗体滴度达到高峰．第28天开始下降。猪弓形虫病血清学诊断的最佳时间为感染后14～28天。ELISA比IHAT抗体检出时间早．抗体滴度高．维持阳性抗体滴度的时间长。 相似文献
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间接血凝试验已广泛应用于寄生虫病的诊断。其中Gill(1965)报道了用甲醛化红细胞间接血凝试验检查家兔和大白鼠血清中的伊氏锥虫抗体;Jatker等(1971)报道了骆驼伊氏锥虫间接血凝试验;Verma等(1977)报道了实验感染水牛犊和黄牛犊的间接血凝滴度。胡氏等(1979)报道了用丙酮醛—戊二醛双醛化红细胞间接血凝试验以诊断耕牛锥虫病,取得了较好的结果。笔者在上述基础上进一步试验,初步标化了各种反应要素,并对疫区耕畜血清,以补体结合反应作对照,检测了伊氏锥虫病抗体。现将结果总结如下。 相似文献
6.
用弓形虫代谢分泌抗原制备间接血凝诊断试剂的研究 总被引：5，自引：0，他引：5
用弓形虫代谢分泌抗原(E／SA)致敏绵羊红细胞制备间接血凝诊断试剂，进行人和动物弓形虫病血清抗体检测，并用弓形虫虫体抗原致敏的红细胞间接血凝诊断试剂进行对照试验。结果表明，在弓形虫阴性、阳性血清检测中，弓形虫E／SA间接血凝诊断试剂与弓形虫虫体抗原间接血凝诊断试剂的符合率为100％；在人工感染兔血的检测中，E／SA致敏的间接血凝诊断试剂比虫体抗原致敏的间接血凝诊断试剂的阳性检出时间提前2d，显示弓形虫E／SA间接血凝诊断试剂具有早期诊断和生前诊断的应用价值。 相似文献
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育肥猪群分别在猪瘟(脾淋源)疫苗免疫第2次和第3次后21d,采血检测免疫抗体。结果表明,猪群第2次免疫后的抗体平均滴度在25.71,高于农业部规定的猪瘟抗体正向间接血凝试验抗体效价≥25的合格标准,且群体合格率为82.8%。第3次免疫后,猪瘟的抗体水平达到了28.24,且群体合格率为99.5%,远远高于农业部规定的合格率必须达到≥70%判定群体合格的标准。 相似文献
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1．1猪瘟采用猪瘟正向间接血凝试验(IHA)检测猪瘟免疫抗体水平，试剂由中国农科院兰州兽医所提供，抗体滴度在1：16以上为阳性。 相似文献
11.
An indirect fluorescent antibody test for the detection of antibody to swine infertility and respiratory syndrome virus in swine sera. 总被引：27，自引：0，他引：27
I J Yoon H S Joo W T Christianson H S Kim J E Collins R B Morrison G D Dial 《Journal of veterinary diagnostic investigation》1992,4(2):144-147
An indirect fluorescent antibody (IFA) test was developed and standardized to detect and quantitate antibody for swine infertility and respiratory syndrome (SIRS) virus in swine sera. Test results were evaluated using sera of pigs infected both experimentally and naturally with SIRS virus. The IFA test used swine alveolar macrophage (SAM) monolayers prepared in 96-well microplates and infected with SIRS virus. The monolayers were incubated with test sera, washed, and stained with fluorescein isothiocyanate-labeled rabbit anti-swine IgG. After another wash step, the monolayers were examined under a fluorescent microscope. A noninfected SAM control well was included for each sample. The antibody titers for each serum sample were recorded as the highest serum dilutions with specific cytoplasmic fluorescence but no fluorescence in the control wells. To evaluate the test, sera of 4 6-week-old pigs that had been infected with SIRS virus, 2 contact pigs, and 13 experimentally infected sows were used. In the experimentally infected pigs, antibody was first detected at 7 days postexposure (PE) and peaked (1:256-1,024) between 11 and 21 days PE. All 13 sow sera were negative at time of infection but were positive (1:64- greater than or equal to 1:1,024) at 14-26 days PE. Seven hundred twenty sera collected from 25 different swine farms with or without a history of SIRS were also tested. Of 344 sera from 15 swine farms with a clinical history of SIRS, 257 (74.7%) sera had IFA titers greater than or equal to 1:4, whereas 371 (98.7%) of 376 sera from herds with no history of SIRS were negative.(ABSTRACT TRUNCATED AT 250 WORDS) 相似文献
12.
WANG Yuchen TIAN Guangyuan ZHOU Yaping GUO Ting ZHAO Hongmei SUN Yajie ZHAO Fengmiao BIAN Yuchen YU Jialiang HAO Yongqing 《中国畜牧兽医》2022,49(8):3122-3130
【Objective】 This study was aimed to verify whether the NP protein of Bovine parainfluenza virus type 3 (BPIV3) could enhance the immune effect of BPIV3 inactivated vaccine【Method】 The antigenicity of the protein encoded by NP gene was analyzed by bioinformatics softwares,and the antigenic region was screened.The truncated NP gene sequence of BPIV3 was amplified by PCR and connected to pET-32a(+) plasmid.Then the high-purity BPIV3 NP protein was obtained by E.coli prokaryotic expression system and Ni affinity chromatography.It was confirmed by Western blotting.BPIV3 was inactivated with 0.3% formaldehyde and mixed with Freund's adjuvant 1:1 to prepare inactivated vaccine.Eight New Zealand White rabbits were randomly divided into four groups with two rabbits in each group,including inactivated vaccine group,NP protein group,inactivated vaccine and NP protein mixed group and control group.Blood samples were collected before and every 7 days after immunization.The levels of specific antibodies and neutralizing antibodies in New Zealand White rabbits of the four groups were measured and compared by indirect ELISA and virus neutralization test.【Result】 DNAStar analysis showed that the average antigen index of amino acid region 193-368 of NP protein was 0.4-1.7,and the hydrophilic index was 0-1.5,which proved that this region had strong antigenicity and hydrophilicity.The NP gene was amplified by PCR and the recombinant expression vector was constructed.Gene sequencing showed that the recombinant expression vector was consistent with the expected results.The results of SDS-PAGE showed that NP protein was highly expressed with a molecular weight of 50 ku and expressed in the form of inclusion body.Western blotting showed that the expressed protein had strong reactivity.The results of ELISA showed that 28 days after immunization,the specific antibody titer of the control group was 0,and the specific antibody titers of inactivated vaccine group,NP protein group and inactivated vaccine and NP protein mixed group reached 1:211,1:217 and 1:218,respectively.The results of virus neutralization test showed that 28 days after immunization,the neutralizing antibody titer of the control group was 0,and the neutralizing antibody titers of inactivated vaccine group,NP protein group and inactivated vaccine and NP protein mixed group were 1:23.32,1:24.48 and 1:24.98,respectively.【Conclusion】 BPIV3 NP protein could enhance the immune effect of BPIV3 inactivated vaccine.Adding NP protein to the inactivated vaccine could be used as a new vaccination method of BPIV3 inactivated vaccine. 相似文献
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A commercial reovirus vaccine alone or experimental reovirus vaccine plus antibody complex were inoculated into 18-day-old specific pathogen free (SPF) broiler embryos at 0.1 of the recommended chick dose. The following groups were used: group 1A was not vaccinated or challenged; group 1B was not vaccinated, but was challenged with virulent reovirus; group 2 received the vaccine complexed with 1/4 dilution of antiserum; group 3 received the vaccine with 1/8 dilution of antiserum; group 4 received the vaccine with 1/16 dilution of antiserum, and group 5 received vaccine alone. At 1, 3, 6, 9, and 12 days of age, serum was collected and antibody against avian reovirus was analyzed by enzyme-linked immunosorbent assay (ELISA). At the same times, spleens were collected and vaccine virus detected by inoculating chicken embryo fibroblasts (CEFs) and examining for cytopathic effect. At 15 days of age, chickens in groups 2-5 were challenged with reovirus. At 22 days of age, birds were euthanatized and weighed. Efficacy of the vaccines was based on safety, percent protection, and antibody response. In ovo vaccination with the commercial or experimental vaccines did not adversely affect hatchability of SPF chickens. The vaccine complexed with antibody resulted in significantly less posthatch mortality (3.7%) when compared to mortality of chickens that received vaccine alone (17%). Both vaccine virus recovery and antibody response were delayed at least 3 days in birds receiving the experimental vaccines. In evo administration of reovirus antibody complex vaccines provided at least 70% protection. The experimental reovirus-antibody complex vaccines were safe and efficacious when given in ovo to SPF broiler embryos. 相似文献
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重组白细胞介素-2提高PRRS抗体阳性猪的猪瘟疫苗免疫效果试验 总被引：6，自引：2，他引：6
观察了重组白细胞介素-2（IL-2）对健康成年猪和PRRS抗体阳性猪的猪瘟疫苗免疫效果的影响。结果显示,重组IL-2和猪瘟疫苗一起免疫健康猪,20d后间接血凝抗体滴度达到1∶64的猪的比例为87.5％,而不注射IL-2的对照组抗体滴度可以达到这一水平的比例只有25％。给经2次猪瘟疫苗免疫但抗体滴度在1∶32以下的PRRS抗体阳性猪单独注射IL-2,20d后,注射前检测不到抗体的猪都检测到了抗体,注射前抗体滴度在1∶8～1∶16之间的猪的抗体滴度提高到1∶32～1∶64。再次用猪瘟疫苗和IL-2共同免疫,可使抗体滴度提高4倍以上。而不注射的对照组抗体滴度则略有下降。说明重组IL-2可以减轻PRRS感染所引起的免疫抑制,提高猪瘟疫苗的免疫效果。 相似文献
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大肠杆菌油佐剂灭活苗免疫雏鸡抗体消长规律的研究 总被引：3，自引：0，他引：3
本试验用鸡源致病性大肠埃希氏菌Ｏ２血清型制成的油佐剂灭活苗对１５日龄雏鸡进行免疫接种，以测定其抗体消长规律；对免疫后不同抗体滴度的雏鸡进行了攻毒试验．结果表明，免疫１周后可监出抗体，２～３周抗体达到高峰，抗体可持续１０周左右．抵抗同型大肠杆菌攻霉的最低抗体滴度为１：１６～１：３．免疫保护期可达２个目以上． 相似文献
16.
An analysis of cellular proliferation, and synthesis of lymphokines and specific antibody in vitro by leucocytes from immunized cattle 总被引：1，自引：0，他引：1
The relationships between the production of lymphokines, cellular proliferation and antibody synthesis by immune bovine PBL in vitro was examined to identify the cellular reactions responsible for differences in the titres of serum antibodies in calves from selected sire lines and MHC Class I phenotypes. Leucocytes from 22 calves immunized with ovalbumin and DNP-BSA proliferated specifically in vitro in the presence of 1-10 micrograms/ml ovalbumin 7-28 days after the second vaccination. Significant correlations between the production of IL-2, IFN-gamma and maximum proliferation were observed for the total group. The quantity of specific antibody produced when PBL were incubated alone or with 10(-1)-10(-2) micrograms/ml ovalbumin was also correlated significantly with the maximum proliferation and the serum antibody titre between 7 and 14 days. Anti-ovalbumin IgG was also synthesized in MLRs where the quantity of antibody was significantly correlated with the magnitude of proliferation. The responses in vitro to DNP-BSA were too low to provide meaningful comparisons. The results indicate that at intervals during the period of increasing serum titres, events in the bovine antibody response in vivo can be replicated in vitro. In addition, assays for proliferation, IL-2 or gamma-IFN, or specific antibody can be used to assess the magnitude of the immune response in vivo in experimental groups of cattle. Significant sire line differences in the serological responses to ovalbumin were observed but DNP-BSA was a poorer antigen and differences in the responses to this antigen were not significant. However, the sire line differences in vivo were not reflected in vitro in proliferative and lymphokine assays; only the production of antibody in vitro was significantly correlated with the in vivo serum titre. 相似文献
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L G Filion H Bielefeldt Ohmann P W Owen L A Babiuk 《Veterinary immunology and immunopathology》1984,7(1):19-32
In the present report, the bovine secondary in vitro antibody response to keyhole limpet hemocyanin is described. The induction of anti-KLH antibody was not dependent upon the presence of mitogen but was antigen specific (KLH vs ovalbumin). Furthermore, this response was dependent upon cell density (10(6) per well), antigen dose (1 to 10(-5) ug/culture) and time in culture (5 days). The antibody produced was specific for KLH as measured in several binding assays. An unresponsive state was detected with high concentrations of KLH (more than 10 ug per culture) which was not due to the formation of immune complexes but to the inactivation of B and/or T cells. The induction of the antibody response was dependent on the presence of macrophages (syngeneic or allogeneic) and their presence could not be replaced with 2-mercaptoethanol. 相似文献
18.
F Petitjean J G Guillet B Vray A D Strosberg J Hoebeke 《Comparative immunology, microbiology and infectious diseases》1987,10(1):9-23
A monoclonal antibody was used to characterize a serogroup 1 specific Legionella pneumophila Philadelphia strain 1 antigenic determinant. A quantitative fluorometric assay was developed to quantitate the antibody sites (2.7 +/- 0.4 X 10(5)) on Legionella bacteria and to determine the physico-chemical parameters of the antibody-antigen interaction (at 4 degrees C: delta G = -10.9 Kcal X mol-1, delta H = 1.7 Kcal X mol-1, delta S = 45 cal X K-1 X mol-1). The same method was used to study the modification or the removal of the antigen by chemical and enzymatic means (trypsin, papain, lysozyme, acetone, chloroform-methanol and Tris-EDTA); only Tris-EDTA extraction resulted in a significant decrease in antibody binding sites. Inhibition studies of the fluorescein-labelled antibody binding were performed with different sugars of which only L-fucosylamine was inhibitory, and with other monoclonal antibodies to Legionella pneumophila serogroup 1 in order to compare their fine specificity and affinity. The results indicate that the epitope recognized was an immunodominant carbohydrate including an aminodideoxyhexose and carried by the lipopolysaccharide. 相似文献
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Muñoz-Zanzi CA Thurmond MC Johnson WO Hietala SK 《Journal of the American Veterinary Medical Association》2002,221(5):678-685
OBJECTIVE: To develop models that could be used to predict, for dairy calves, the age at which colostrum-derived bovine viral diarrhea virus (BVDV) antibodies would no longer offer protection against infection or interfere with vaccination. DESIGN: Prospective observational field study. ANIMALS: 466 calves in 2 California dairy herds. PROCEDURE: Serum BVDV neutralizing antibody titers were measured from birth through 300 days of age. The age by which colostrum-derived BVDV antibodies had decayed sufficiently that calves were considered susceptible to BVDV infection (ie, titer < or = 1:16) or calves became seronegative was modeled with survival analysis methods. Mixed-effects regression analysis was used to model colostrum-derived BVDV antibody titer for any given age. RESULTS: Half the calves in both herds became seronegative for BVDV type I by 141 days of age and for BVDV type II by 114 days of age. Rate of antibody decay was significantly associated with antibody titer at 1 to 3 days of age and with whether calves were congenitally infected with BVDV. Three-month-old calves were predicted to have a mean BVDV type-I antibody titer of 1:32 and a mean BVDV type-II antibody titer of 1:16. CONCLUSIONS AND CLINICAL RELEVANCE: Results provide an improved understanding of the decay of BVDV-specific colostrum-derived antibodies in dairy calves raised under typical field conditions. Knowledge of the age when the calf herd becomes susceptible can be useful when designing vaccination programs aimed at minimizing negative effects of colostrum-derived antibodies on vaccine efficacy while maximizing overall calf herd immunity. 相似文献
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试验旨在筛选并制备鸡PD-1单克隆抗体,对该单克隆抗体的免疫学特性、结合活性及其对鸡PD-1/PD-L1信号通路激活的阻断作用进行初步研究。运用杂交瘤细胞融合技术筛选杂交瘤细胞株,采用ELISA方法、Ig抗体亚型鉴定试剂盒、Western blotting鉴定抗体的免疫学特性,利用间接免疫荧光技术及流式细胞术检测筛选单抗与鸡PBMCs的结合情况,应用该单抗与IBDV感染7 d后的鸡PBMC细胞作用,利用实时荧光定量PCR技术检测IL-2、IL-6和IFN-γ等细胞因子的表达情况。结果显示,试验获得1株特异、稳定分泌鸡PD-1单克隆抗体的杂交瘤细胞株,命名为PD-1-D05。该单克隆抗体的亚型属于IgG1,杂交瘤细胞培养上清和腹水的效价分别为1：211和1：2.048×105。ELISA和Western blotting检测结果表明,PD-1-D05单抗能与免疫原发生特异性反应,与pET-28a （+）、Rosetta菌株蛋白提取液上清及无关蛋白无交叉反应。间接免疫荧光及流式细胞术检测结果显示,PD-1-D05单克隆抗体能与鸡PBMC特异性结合,且IBDV感染7 d后的PBMC经单抗处理后,IL-2表达量显著升高（P<0.05）,IFN-γ转录水平显著下降（P<0.05）,IL-6表达水平较IBDV攻毒组细胞虽有所下降,但并无统计学差异（P>0.05）。结果表明,试验成功筛选并制备了能够稳定分泌鸡PD-1单克隆抗体的细胞株,所获得的PD-1单克隆抗体具有良好的免疫学特性,该单抗能够特异性识别鸡PD-1分子并与鸡PBMC细胞特异结合,并在一定程度上恢复由于IBDV感染导致的PD-1/PD-L1信号通路激活引发的免疫调节相关细胞因子IL-2、IFN-γ的异常表达。 相似文献