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1.
本研究应用间接ELISA方法对新城疫LaSota和V4疫苗免疫SPF鸡及免疫后攻毒SPF鸡的血清中新城疫病毒（NDV）特异性HI抗体、IgM和IgG抗体水平的动态变化进行了检测。结果表明，V4较LaSota疫苗免疫鸡HI抗体提前3天左右出现，但除高峰期（1周左右）外，其HI抗体水平均低于LaSota免疫鸡。LaSota免疫鸡。LaSota疫羁免疫鸡血清中NDV特异性IgM和IgGr抗体高峰较V4免疫鸡提前约2周出现，攻毒后，LaSota免疫鸡血清中特异性IgG和IgG回忆应答显著，而V4免疫鸡IgM和IgG回忆应答不明显。  相似文献   

2.
为了观察小鼠感染不同种旋毛虫后肉汁抗体水平的变化及其与血清抗体水平的相关性,将200只昆明小鼠随机分成4组（每组50只）,每只分别感染300条乡土旋毛虫（T2）、布氏旋毛虫（T3）、伪旋毛虫（T4）及纳氏旋毛虫（T7）幼虫,感染后2～6周每组每周随机剖杀10只小鼠,收集血清及肉汁,用旋毛虫肌幼虫ES抗原ELISA检测血清及肉汁中抗体水平;另取40只昆明小鼠随机分成4组（每组10只）,每只分别感染500条T2、T3、T4、T7幼虫,感染后6周剖杀,制备肉样,用ELISA检测4℃及-20℃保存不同时间后的肉汁抗旋毛虫抗体动态。T2、T3、T4、T7感染小鼠后的肉汁抗体水平和变化趋势相似,均是在感染后第3周检测出肉汁特异性抗体,抗体阳性率分别为60％、30％、30％、30％,至感染后第4周肉汁抗体阳性率均升至100％。4组小鼠感染后2～6周的肉汁与血清抗体水平均具有相关性。T2、T3、T4、T7感染小鼠肌肉4℃保存7d与1d的肉汁抗体水平相比均无显著性差异;所有的感染小鼠肌肉-20℃保存2个月的肉汁抗体水平与保存1个月的相比均无显著性差异;虽然保存3个月的肉汁抗体水平与保存1个月的相比均已有显著性差异,但抗体阳性率仍均为100％并持续至实验结束时的4个月保存期。结果表明,肉汁中抗旋毛虫抗体的检测可用于新鲜肉、冷藏肉及冷冻肉中乡土旋毛虫、布氏旋毛虫、伪旋毛虫、纳氏旋毛虫捡疫的初筛。  相似文献   

3.
东方田鼠抗日本血吸虫抗体水平检测与分析   总被引：2，自引：0，他引：2  
东方田鼠具有抗日本血吸虫病的特性，观察东方田鼠感染日本血吸虫尾蚴后的特异性抗体水平变化，可为阐明其抗病机制提供依据。本试验用间接ELISA方法检测东方田鼠抗成虫可溶性抗原（SWAP）和几种日本血吸虫基因重组抗原（Sj28-GST、LHD-SjTP1、C-SjParamyosion）的特异性抗体水平动态变化，并检测东方田鼠感染日本血吸虫尾蚴前、后的IgG3亚类特异性抗体水平。结果东方田鼠在攻击日本血吸虫尾蚴后，抗SWAP的特异性抗体水平有所升高，而抗几体基因重组抗原的特异性抗体水平低，没有显著变化。东方田鼠正常血清中抗SWAP和IgG3亚类抗体的OD值是牛血清白蛋白（BSA）对照的31倍。结果表明，在东方田鼠血清中有较高水平的SWAP的IgG3亚类抗体，值得进一步深入探讨。  相似文献   

4.
为探讨猪繁殖与呼吸综合征病毒(PRRSV)GP5和M蛋白滴鼻免疫小鼠后产生的特异性抗体的中和活性,研究以原核表达的重组蛋白GP5和M为抗原分别与霍乱毒素B亚基(CTB)佐剂按10∶1的比例混合制备成CTB黏膜佐剂疫苗,滴鼻免疫小鼠,于首次免疫后0,7,14,21,28,35,42 d收集唾液和血清样品,利用间接ELISA方法分别检测免疫小鼠唾液中特异性抗体IgA,结果均于28 d时S/N值达到最高,ELISA效价均为1∶2;血清中特异性抗体IgG分别于21,28 d时S/N值达到最高,ELISA效价均为1∶800。通过本实验室建立的基于PAM细胞中和试验方法分别检测唾液中特异性抗体IgA和血清中特异性抗体IgG中和滴度均1∶2,结果表明PRRSV GP5和M蛋白滴鼻免疫小鼠后产生的特异性抗体未达到免疫保护作用的中和抗体水平。  相似文献   

5.
鱼精蛋白增强抗原诱导早期体液免疫反应的研究   总被引：1，自引：0，他引：1  
为探究鱼精蛋白(PP)对抗原诱导早期体液免疫反应的增强作用,本研究采用PP与鸡卵清白蛋白(OVA)联合免疫小鼠,利用ELISA方法检测了免疫后10d内小鼠血清特异性抗体、细胞因子表达变化水平,采用流式细胞术检测了PP对外周血CD4~+和CD8~+T细胞亚群的影响,并进行了免疫小鼠的攻毒保护试验。结果显示,PP能快速激活OVA诱导的特异性抗体(IgM、IgG)应答,促进Th2偏向的免疫反应(IgG1、IL-5、IL-10),而对Th1型免疫应答(IgG2a、IFN-γ)无显著影响。免疫后10d的外周血淋巴细胞流式细胞术检测结果显示,PP显著提高外周血CD4~+/CD8~+值。此外,PP作为佐剂显著提高了大肠杆菌疫苗免疫小鼠3d后的攻毒存活率,增强了大肠杆菌疫苗的早期免疫保护。本研究为PP作为候选疫苗佐剂及其临床应用潜力提供了实验依据。  相似文献   

6.
以日本血吸虫基因重组抗原LHD-Sj23／pGEX、可溶性虫卵抗原（soluble egg antigen，SEA）及SEA经Sephadex G-200柱层析分离后得到的第一峰SEAl作为诊断抗原，应用ELISA检测人工感染日本血吸虫绵羊血清及自然感染日本血吸虫水牛血清。结果显示，三种抗原用于检测人工感染日本血吸虫绵羊血清的阳性符合率分剐为88．79％、98．15％、100％，阴性符合率均为100％；自然感染日本血吸虫水牛血清的阳性符合率分别为80％、80％、84％，阴性符合率分别为88．9％、93．3％、91．1％；三种抗原与伊氏锥虫病牛血清无交叉反应；检测感染日本血吸虫不同时间绵羊血清，发现感染6周后均被检出针对这三种抗原的特异性抗体，且SEA及SEAl在感染4周后即可检出特异性抗体。三种抗原的诊断效果差异不显著。  相似文献   

7.
将制备的狂犬病rSRV9减毒口服冻干脂质体活疫苗配合以自制的复合佐剂,免疫小鼠、犬、猫,根据ELISA抗体定量检测方法,对免疫后小鼠血清IgG、小鼠粪便IgA、犬和猫血清IgG、犬和猫唾液IgA的ELISA抗体水平进行检测,其检测结果与注射狂犬病疫苗免疫效果进行对比。结果显示,复合佐剂+rSRV9病毒口服免疫后70d,小鼠血清抗狂犬病特异性IgG抗体水平为4 214.00U/mL,小鼠粪便中SIgA抗体水平为137.00U/mL;复合佐剂+rSRV9病毒口服免疫犬120d抗体水平最高,犬抗狂犬病特异性IgG抗体水平为1 420.00U/mL,唾液中SIgA抗体水平为1 199.00U/mL;猫抗狂犬病特异性IgG抗体水平为2 088.00U/mL,唾液中SIgA抗体水平为1 896.00U/mL。狂犬病疫苗口服组与注射组特异性抗体水平差异均不显著。结果表明,狂犬病rSRV9减毒口服冻干脂质体活疫苗的抗体产生水平已接近注射狂犬病疫苗的免疫效果,能够达到对动物的免疫效果。  相似文献   

8.
应用酶联免疫吸附试验(ELISA)检测猪传染性胃肠炎患猪不同时期血清中IgG、IgM的变化规律,结果显示:感染后15d特异性IgG与IgM的阳性率分别为34.55%(19/55)和78.18%(43/55);感染后45d特异性IgG与IgM的阳性率分别为92.73%(51/55)和47.27%(26/55)。通过检测,不仅可为临床鉴别猪流行性腹泻(PED)与猪轮状病毒感染提供实验依据,而且还表明猪感染TGEV后的体液免疫状况和抗体产生水平及其变化规律。  相似文献   

9.
以纯化的金黄色葡萄球菌无毒诱变超抗原GST-mTSST-1融合蛋白作为检测抗原，通过优化ELISA反应条件，初步建立了检测血清特异性GST-mTSST-1抗体的间接ELISA方法，监测了小鼠血清中特异性GST-mTSST-1抗体水平的动态变化规律。方阵试验确定的GST-mTSST-1抗原的最适包被浓度为5．0μg／mL，血清最佳稀释倍数为1：100，ELISA阳性反应的临界值为D490nm≥0．219，批内和批间重复试验的变异系数均小于10％。应用该方法对试验小鼠血清进行检测，结果表明，建立的间接ELISA方法具有较高的敏感性、特异性和较好的重复性，适用于检测特异性GST-mTSST-1血清抗体。  相似文献   

10.
将3种检测口蹄疫非结构蛋白抗体的间接ELISA试验进行了比较。这3种间接ELISA试验检测田间血清样品的最低符合率为96％，最高符合率达98％：有2种间接ELISA试验可以检测到感染口蹄疫第10天后血清中的口蹄疫感染抗体．有1种间接ELISA试验可以检测到感染口蹄疫第14天后血清中的口蹄疫感染抗体；这3种间接ELISA试验检测12份已知口蹄疫阳性血清的试验结果吻合。研究表明．这3种间接ELISA试验方法对于检测口蹄疫感染抗体具有较好的特异性．其中猪口蹄疫3A蛋白间接ELISA诊断试剂盒在灵敏性、特异性和符合率方面更优于另外2种试剂盒。  相似文献   

11.
An IgM enzyme-linked immunosorbent assay (IgM-ELISA) for the detection of antibodies to bovine herpesvirus-1 (BHV-1) was developed. Its applicability was examined by serological studies in two calves experimentally infected with virulent BHV-1 over a period of 60 days. IgM antibodies were detected by ELISA on day 6 after infection, and there was an increase in IgG antibodies on day 9. Serum neutralizing (SN) antibodies were detected only on day 13, confirming the higher sensitivity of the ELISAs. A similar study of four calves treated with a commercial inactivated virus vaccine indicated no detectable IgM-ELISA response, and late SN and IgG-ELISA reactivity. Thus IgM-ELISA appears to be of value in assessing recent infection, whereas IgG-ELISA and SN cannot distinguish between infection and vaccination. The possible limitations imposed on the specific IgM-ELISA by the presence of IgM rheumatoid factor (IgM-RF) in bovine serum were examined. IgM-RF levels were determined in bovines of various ages. Elevated values of IgM-RF were found in the sera of older animals; their occurrence may lead to false IgM-positive diagnosis (16%) of BHV-1 infection. This was examined in 113 serum specimens from suspected cases of BHV-1 infection and in 32 bulls at an insemination center. Pretreatment of serum samples with an antibovine IgG serum eliminated false positivity of the IgM-ELISA. It is concluded that IgM-ELISA should be of particular value in the diagnosis of recent infection with BHV-1, mainly in calves.  相似文献   

12.
An antibody-capture enzyme-linked immunosorbent assay (ELISA) for detection of immunoglobulin (Ig) M antibodies to bovine respiratory syncytial virus (BRSV) in cattle was developed. Monoclonal antibody to bovine IgM was used as the catching antibody. The IgM-ELISA was used, as well as a BRSV-specific IgG ELISA to determine the kinetics of IgM and IgG antibody responses to BRSV infections in cattle. High IgM and IgG antibody titers developed after naturally occurring or induced BRSV infection of calves (6 to 7 months old). Induced infection resulted in an IgM response that was first detectable at postinoculation day (PID) 11 reached a maximum at PID 13, and became undetectable again about PID 28. An IgG response also was detected by PID 11. However, a maximum response was not reached before PID 23, and titers remained high (until PID 80). In naturally occurring infection, IgM and IgG responses in calves were observed in the acute phase of epizootics of respiratory tract disease. Patterns of IgM and IgG response curves were similar to those observed in experimentally infected calves. The involvement of BRSV in an epizootic of respiratory tract disease in 8 calves (2 to 3 weeks old) was demonstrated by the detection of BRSV in several lung lavage samples. All calves had existing IgG antibodies to BRSV which were interpreted to be maternally derived. None of the calves responded with an increase in IgG antibody titer. However, a weak but distinct BRSV IgM antibody response occurred in 6 calves.(ABSTRACT TRUNCATED AT 250 WORDS)  相似文献   

13.
Goats housed in microbiologically secure facilities were experimentally endobronchially infected with Mycoplasma capricolum subsp. capripneumoniae (Mccp), causal agent of contagious caprine pleuropneumonia (CCPP). The animals were monitored over an 8-week period post-infection (p.i.). Elevated temperatures were observed 2-7 days p.i., reaching a maximum of 41.5 degrees C in one animal (1884). By 8 weeks p.i. the infection was successfully cleared, with no Mccp being recovered from the lungs, serum or nasal passages. Mccp was not isolated from serum throughout the experiment, either directly by culture or indirectly via polymerase chain reaction (PCR). Humoral immune responses against Mccp capsular polysaccharide (CPS) were generally poor when measured by ELISA. CPS antigen was present in the serum of all infected animals early in the infection (day 14 p.i.), although in one animal (1855) CPS antigen persisted throughout. This was the only animal to exhibit a serious cough (day 5-19 p.i.). Successful diagnosis of CCPP was achieved using two different types of latex agglutination test (CPS antibody and CPS antigen detection test), immunoblotting and a blocking ELISA, although the latter lacked sensitivity until later in the infection (35-40 days p.i.). Only a single animal (1855) was detected positive using the current complement fixation test (CFT). Strong immune responses to protein antigens were detected by IgG and IgM immunoblotting from the first time point at day 14 p.i. IgM immunodominant bands of 220, 85, 62 and 40kDa were observed in the 3 infected animals and from CFT-positive CCPP field sera. Band intensity gradually diminished throughout the experiment. IgG immunodominant bands of 108, 70, 62, 44, 40 and 23kDa were shared between experimentally-infected and field sera, with band intensity either remaining unchanged or increasing from day 14 p.i. These bands were not present using pre-infection sera. Of the diagnostic tests used, only the CPS antibody detection latex agglutination test and IgG immunoblotting gave positive diagnoses throughout the entire period post-infection (days 14-53 p.i.).  相似文献   

14.
Antibody (immunoglobulin G (IgG), IgM or IgA) levels relative to ferritin in six foal sera (three male and three female) after birth (day 0 and 2, 6, 10, 20, 28, 36, 40, 52 and 56 weeks of age) were semi‐quantitatively measured with normalization with antibody activity to ferritin in one adult horse serum. After addition of horse spleen ferritin to the serum sample, the complex formed between antibodies to ferritin in the serum and ferritin was co‐immunoprecipitated using antibody to horse spleen ferritin. Antibody classes of the co‐immnoprecipitate were detected with antibodies specific for horse IgG, IgM or IgA heavy chain. Six adult horse serum samples were found to have ferritin‐binding activities in all immunoglobulin classes examined. Although ferritin antibody activities (IgG, IgM and IgA) were scant in the foal sera before sucking colostrum (day 0), their activities increased at 2 weeks of age. IgG antibodies showed a biphasic response and IgM antibody activity increased up to 40 weeks of age. Antibody (IgG, IgM and IgA) activities to ferritin in three colostrum samples were significantly higher than in adult horse serum samples. These results demonstrate that antibody to ferritin in foal serum is derived from colostrum after birth and is produced thereafter.  相似文献   

15.
A quantitative competitive binding "triple-sandwich" enzyme immunoassay was developed and used to evaluated pathogen/class-specific antibody responses in Holstein-Friesian cows vaccinated against Clostridium perfringens B-toxin. Vaccination of cows at six weeks and again at two weeks prepartum increased pathogen-specific IgG levels in each dam's colostrum and respective calf's serum. Pathogen-specific IgG and IgM concentrations in dams' sera and colostra were related to subsequent pathogen-specific IgG and IgM neonatal sera concentrations. Only pathogen-specific IgA in dams' colostra was correlated to neonatal levels, possibly owing to a different origin and role of this immunoglobulin class. All class-specific colostral immunoglobulin levels were related to subsequent neonatal concentrations. Isotypic antibody responses against C. perfringens B-toxin were found with pathogen-specific IgM predominant in dams' sera and pathogen-specific IgA predominant in colostra and neonatal sera.  相似文献   

16.
SUMMARY: A factorial-design survey was performed to determine the prevalence of specific antibody against Toxoplasma in young and adult sheep from 6 areas in 3 different geoclimatic zones in South Australia. Serum samples obtained from 1,159 sheep belonging to 59 flocks were tested by a conventional indirect haemagglutination test (IHAT) as well as by an enzyme-linked immunosorbent assay (ELISA) which used class-specific conjugates to detect both IgG and IgM against Toxoplasma. Titres > 64 were detected In 7.4%, 9.2% and 25.2% of the sheep by the IHAT, IgG-ELISA and IgM-ELISA respectively. A significant positive correlation was found between the results of the IgG-ELISA and the IHAT with identical results obtained for 1,050 samples. Antibody detected by all 3 tests was more prevalent and higher in titre in adult sheep than in lambs as well as in sheep from Kangaroo Island than in those from mainland South Australia. Although the regional differences in prevalence suggested a relationship with climate, no significant correlations were detected between the prevalence results and any single climatic factor; namely, average annual rainfall, average annual evaporation or mean temperature range.  相似文献   

17.
The antibody response of 20 pregnant ewes to oocyst infection with Toxoplasma gondii was determined by the latex agglutination test (LAT) and compared with the indirect fluorescent antibody test (IFAT) and a commercially available indirect haemagglutination test (IHAT). The LAT and IFAT showed a similar rapid response with antibody first appearing by two to three weeks after infection and titres that correlated closely (r = 0.81, P less than 0.001). The IHAT response was slower and less consistent up to seven weeks after infection. The LAT response was biphasic in seven of the sheep. Sera were fractionated using a minicolumn gel filtration technique and specific IgM and IgG titres determined by LAT. IgM titres peaked three weeks after infection and IgG titres exceeded IgM titres at a mean time of 4.7 weeks after infection (range 3 to 7). Eleven sheep exhibited fetopathy with abortion/parturition 12 to 53 days after infection; in nine of them IgG titres exceeded IgM at that time. A non-specific anti-toxoplasma reaction associated with IgM antibody occurred at low titre in one sheep. The results indicate that used from a dilution of 1/64 the LAT is a sensitive, reliable and rapidly responsive serological test for toxoplasma infection in ewes and it may be utilised with sample fractionation techniques to determine IgM titres. It is suggested that the best time to examine ewe sera to assist diagnosis of toxoplasma abortion is one week after abortion. While the determination of specific IgM titres in ewe sera may assist epidemiological studies and, sometimes, diagnosis, in the majority of aborting ewe sera it is unlikely to aid diagnosis.  相似文献   

18.
The diagnosis of acute babesiosis by direct examination of blood smears has some limitations and the indirect serological methods currently in use are designed for detection of IgG, which may not be detectable at an early stage of infection. There is a need, therefore, for rapid and reliable procedures to diagnose acute infections. An ELISA system using a crude antigenic preparation of Babesia bovis was standardized for the detection of IgM antibodies. Optimal dilutions of the antigen, using positive and negative reference sera, were determined by checkerboard titrations. Serum samples of cattle imported from tick-free areas collected before and during an immunization process were used to validate the tests. The specificity was 94% and sensitivity 100%. Specific IgM antibodies against B. bovis first appeared on the 11th day post-inoculation (p.i.) in animals infested with Boophilus microplus ticks and on the 19th day p.i. in animals which had been inoculated with infected blood. Antibody titers decreased after Day 33; however, all animals remained positive until the end of the experiment (124 days).  相似文献   

19.
An IgM capture ELISA using heterologous antibodies was developed to evaluate the kinetics of the humoral immune response in dogs experimentally infected with Toxoplasma gondii RH strain. Detection of parasite in tissues from inoculated dogs was evaluated by mouse bioassay and immunohistochemical techniques. Serum samples were obtained at regular intervals up to 62 days post-inoculation (p.i.), when the animals were necropsied and their tissues examined. Antibody levels were measured by IgM capture ELISA (McELISA), indirect hemagglutination (IHA), indirect fluorescent antibody test (IgG-IFAT) and indirect immunoenzymatic assay (IgG-ELISA). All dogs seroconverted but only one exhibited severe clinical signs of infection. IgM antibodies were detected by McELISA from the seventh day on, with decreasing IgM levels around the 27th day. Similar results were obtained from IHA, although McELISA showed earlier and longer detection of IgM antibodies. IgG antibodies were detected from the seventh day on, and throughout the period of observation. Immunohistochemical findings and mouse bioassay revealed the presence of free tachyzoites in tissues of the clinically affected dog only. These results suggest that T. gondii acute infection in dogs shows a remarkably transient IgM synthesis, and this feature may constitute an important marker of active infection. Furthermore, McELISA was shown to be a potential tool to diagnose canine toxoplasmosis.  相似文献   

20.
An experimental reproduction of CBPP was implemented and the animals were surveyed for serology during 4 months. The ELISA/IgG test detects the antibodies few days after the CF test but is more precise for detection on the longer term. The early antibody detection can be done with the ELISA/IgM test. Circulating antigen (galactan) has been detected in a cow that died of an acute form of CBPP. Excretion of mycoplasmas starts 1 to 2 weeks before the seroconversion: the ELISA/IgG test remains positive during the excretion phase and even longer.  相似文献   

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