Comparison of the enzyme-linked immunosorbent assay and the indirect hemagglutination and complement fixation tests for detecting antibodies to Mycoplasma hyopneumoniae.

Caesarean-derived, colostrum-deprived swine were exposed to a broth culture of a low passage field isolate of Mycoplasma hyopneumoniae by intranasal inoculation. The intranasal-inoculated swine subsequently were commingled with their litter-mates to effect transmission via contact-exposure. Sera were collected from the swine at two to four week intervals for approximately one year postexposure and evaluated by the enzyme-linked immunosorbent assay (ELISA), indirect hemagglutination and complement fixation tests. The intranasal-exposed swine seroconverted earlier, developed higher titers and remained indirect hemagglutination and complement fixation positive longer than the contact-exposed swine. It was concluded that the antibody response of intranasal-exposed swine was artificially high and that sera from such swine were not suitable for evaluating the sensitivity of mycoplasmal pneumonia of swine serodiagnostic tests. The indirect hemagglutination test was relatively insensitive and technically cumbersome and the least promising as a practical field test. The complement fixation test appeared to be slightly more sensitive in detecting early antibody production (especially in contact-exposed swine) but it was the least sensitive in detecting late antibodies. The ELISA was generally the most sensitive procedure. Individual high ELISA titers were from ten to 32 times greater than maximum complement fixation and indirect hemagglutination titers. The most striking difference among the three tests was the persistence of high ELISA titers late in the study. All swine were ELISA positive at necropsy approximately one year postexposure despite the fact that lungs were devoid of lesions and culturally and immunofluorescent negative for M. hyopneumoniae.     

上海地区规模化猪场弓形虫病的血清学调查

用正向间接血凝试验对2002年-2007年间从上海地区8个区县的规模化猪场采集1 377份血样进行了猪弓形虫病抗体的检测.通过连续6年对上海地区规模化猪场的监测发现,2002年所有猪场均无弓形虫病感染,从2003年开始出现了较高水平的感染率,最高达到66.7%.随后几年里不同猪场出现了程度不同的感染,2004年和2005年其感染率逐年下降,到2005年达到最低值1.6%,而2006年其感染率发生了反弹,升高到48.7%,2007年又出现了下降的趋势,下降到13.1%.掌握上海地区猪弓形虫病的感染状况,为基层的防治工作提供技术指导.     

Development of a blocking ELISA for screening antibodies to porcine rubulavirus, La Piedad Michoacan Virus.

A blocking enzyme-linked immunosorbent assay (ELISA) was developed to detect antibodies to porcine rubulavirus (La Piedad Michoacan Virus [LPMV]) in serum samples from pigs. The test, based on a monoclonal antibody against the LPMV hemagglutinin-neuraminidase glycoprotein, had a sensitivity of 99% and a specificity of 97%. The results of this test were in agreement with those obtained by an indirect ELISA and hemagglutination inhibition, indirect immunofluorescence, and virus neutralization tests. The blocking ELISA is considered the most suitable test for routine screening for antibodies against LPMV.     

猪圆环病毒间接免疫荧光方法的建立

将接种PCV2且经多次传代的PK-15带毒细胞分散到96孔细胞培养板上，制成抗原诊断板，应用间接免疫荧光技术检测血清中PCV2的抗体。通过对各种反应物的工作浓度与作用时间的优化，建立了PCV2抗体的间接免疫荧光检测技术，可用于PCV2抗体水平的监测，为今后的猪圆环病毒疫苗抗体检测的研究提供了检测方法。     

Serological investigation of horse sera for antibodies against mycoplasmas and acholeplasmas

Sera from horses with respiratory disease (RD) have been investigated using the complement fixation test, indirect hemagglutination test, enzyme immune assay, and the metabolic inhibition test, and sera from mares after abortion, using the complement fixation test, indirect hemagglutination test and enzyme immune assay, for antibodies against Mycoplasma equirhinis, M. subdolum, M. equigenitalium, M. pulmonis, M. felis, Acholeplasma laidlawii, A. hippikon and A. equifetale. Antibodies were found against all mycoplasma and acholeplasma species tested, more often against acholeplasmas. The antibody pattern was quite similar for horses with RD and for mares after abortion. The results of the four serological tests performed showed only a limited correlation and the percentage of sera with antibodies detected by the four tests used differed widely.     

The indirect fluorescent antibody technique as a method for detecting antibodies in aborted fetuses.

In this investigation the indirect fluorescent antibody technique was used to titrate antibodies in bovine sera to parainfluenza 3, infectious bovine rhinotracheitis virus and bovine viral diarrhea virus. These results were compared to those determined on the same samples by hemagglutination inhibition for parainfluenza 3 virus and serum neutralization for bovine virus diarrhea and infectious bovine rhinotracheitis virus. The results of the serological methods agreed closely. The indirect fluorescent antibody technique is a rapid and sensitive method for detecting antibodies and the procedure lends itself to use in diagnostic laboratories. In addition to the above viruses the presence or absence of antibodies to bovine coronavirus and bovine adenovirus 3 were determined by the indirect fluorescent antibody technique in thoracic fluids from 100 aborted fetuses and 50 nonaborted fetuses. Results on these samples were not compared to hemagglutination inhibition or serum neutralization as the condition of fluid samples from aborted fetuses renders interpretation of such tests unreliable. Antibodies to one or more viruses were detected in 30 of the 100 aborted fetuses and in seven of the 50 nonaborted fetuses. Antibodies to more than one agent were detected in eleven of the 100 aborted and in one of the 50 nonaborted fetuses. Reasons for this occurrence and application of the test in determination of causes of abortion are discussed.     

Serological comparison of type strains of porcine, bovine, and ovine mycoplasmas with atypical colony morphology

The type strains of Mycoplasma hyopneumoniae, M. flocculare, M. dispar, and M. ovipneumoniae, all characterized by nipple-less colonies on solid media, were compared serologically. By indirect hemagglutination and by complement fixation tests they were found to constitute a related group. By crossed immunoelectrophoresis a sharing of common antigens was demonstrated, whereas no cross reactivity was noted by the metabolism inhibition test.The type strains of Mycoplasma hyorhinis and Mycoplasma bovirhinis were included in the study for comparison. Although some cross reaction was noted, they appeared just moderately related to the nipple-less group as well as to each other.     

猪胸膜肺炎放线杆菌血清3型间接血凝试验检测方法的建立

本试验用分离纯化的猪胸膜肺炎放线杆菌（APP）3型荚膜多糖（CPS）,致敏双醛化处理的绵羊血红细胞,建立了APP的血清学诊断方法,结果表明,该方法特异性强,仅与APP 3型阳性血清反应,可以用其进行APP疫苗免疫后抗体检测,该诊断方法简单、快速,适用于临床疾病的诊断。     

Immunologic and pathologic responses of cows to naturally occurring Mycoplasma bovis mastitis

Systemic humoral and cell-mediated immune responses of four Holstein cows with natural Mycoplasma bovis mastitis were evaluated to determine whether a relationship exists between systemic cellular and humoral responses and the pathogenesis and resolution of infection. In vitro lymphocyte activation tests of peripheral blood lymphocytes and in vivo skin tests with M. bovis antigens provided evidence that cell-mediated immune responses against M. bovis may be involved in successful resolution or containment of infection. In several observation it appeared that viable M. bovis and their aqueous extracts are suppressive to cell-mediated responses.Humoral responses were determined by the serum indirect hemagglutination (IHA) test and the growth inhibition test. The IHA titers after approximately 2 weeks of infection were elevated; however, 75–87% of the IHA activity was in the IgM antibody class.The cell-mediated immune responses may be necessary for resolution of mycoplasmal mastitis both directly and via their helper cell function on antibody production. However, it appears that immune injury to mammary tissue results from the immunologic response to infective mycoplasma. Presence of locally secreted antibody and locally active immune cells may provide a better indication of those animals in the process of resolving the infection than was observed using systemic indicators of immune responsiveness such as indirect hemagglutination or growth inhibition tests.     

Studies on the antigenicity of an inactivated, aluminum hydroxide adjuvant equine influenza vaccine.

An inactivated, aluminum hydroxide adjuvant equine influenza vaccine was tested in horses and guinea pigs to determine the levels of antigen that would elicit maximum serological responses. Vaccine containing serial twofold increments of A/Equi-1/Prague and A/Equi-2/Miami strains of equine influenza virus was administered to random groupings of both types of test animals. The hemagglutination inhibition antibody response for each group was then measured. Results in horses and guinea pigs were compared to determine if the equine serological values could be related to a potency test in laboratory animals. The highest mean hemagglutination inhibition antibody response in horses occurred in groups vaccinated, respectively, with 128 or 256 hemagglutination units of A/Equi-1 and 512 or 1024 hemagglutination units of A/Equi-2 antigen. Groups vaccinated with further two- or fourfold increases in these antigens had mean hemagglutination inhibition titers that were somewhat lower than the maximum levels. When graded doses of vaccine were given to guinea pigs, their hemagglutination inhibition antibody titers reached a plateau of maximum values, similar to the serological response in vaccinated horses. Test horses remained clinically free from signs of equine influenza during the year following vaccination and no untoward post-vaccination reactions were observed.     

抗牛多杀性巴氏杆菌高免卵黄抗体IgY的制备及间接血凝检测方法的建立

本研究通过自制牛源荚膜血清A型多杀性巴氏杆菌灭活菌苗免疫产蛋鸡,采用卡拉胶结合硫酸铵沉淀法提取卵黄抗体IgY,并采用间接血凝方法检测抗体效价。结果表明,抗原致敏红细胞的最佳浓度是800μg/mL,免疫后第7周抗体效价达到高峰,效价为1：1024,高效价持续5周开始下降,测定提取的IgY浓度为8.258mg/mL,无菌检测及动物安全性实验表明制备的卵黄抗体安全可靠。本研究制备了抗牛多杀性巴氏杆菌卵黄抗体,为防治由荚膜血清A型多杀性巴氏杆菌所致的犊牛肺炎提供了新的手段。     

Indirect hemagglutination test in equine infectious anemia.

An indirect hemagglutination was developed for the diagnosis of equine infectious anemia using sheep red blood cells coated with group specific virus antigen which had been highly purified by affinity chromatography. The presence of indirect hemagglutination antibodies was demonstrated in horses with equine infectious anemia since the cells were specifically agglutinated by all the serum samples obtained from experimentally infected horses. Antibodies appeared within 35 days after inoculation, and development of which coincided well with that of precipitating and complement fixing antibodies. Titer of indirect hemagglutination antibodies were ten to 320 times greater than those of precipitating antibodies. Test results could be read more clearly by the indirect hemagglutination test especially in weakly positive cases. Ninety-six samples from suspected field cases collected from every region of Japan which were positive on the immunodiffusion test were also positive on indirect hemagglutination test. Serum samples from 420 horses in one race track were examined by both the indirect hemagglutination and immunodiffusion tests to determine the reliability of the indirect hemagglutination test for diagnosis of equine infectious anemia. The same result was obtained on both tests. Based on this evidence, the indirect hemagglutination test can be employed as a very sensitive serological test for the diagnosis of equine infectious anemia.     

猪O型口蹄疫免疫抗体检测方法的比较

试验采用口蹄疫O型间接血凝试验和O型口蹄疫抗体液相阻断ELISA 2种试验方法检测了60份血清中猪O型口蹄疫免疫抗体.研究结果显示,60份被检血清口蹄疫O型间接血凝试验检测合格率为93.3％,O型口蹄疫抗体液相阻断ELISA试验检测的合格率为73.3％,口蹄疫O型间接血凝试验检测合格率明显高于O型口蹄疫抗体液相阻断ELISA试验检测的合格率（相差20个百分点）;2种方法的总符合率为66.7％、＜25（＜2^6）的符合率为28.6％,≥2^5（≥2^6）的符合率为82.1％.2种方法检测出的整体免疫效果较好,平均合格率远高于农业部规定的70％.     

猪流行性腹泻病毒实验感染猪血清抗体的消长规律

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。     

Comparison of serological tests for the detection of antibody to natural and experimental murine cytomegalovirus.

Three serological tests, i.e. complement fixation test, indirect immunofluorescent assay, and enzyme-linked immunosorbent assay (ELISA) were compared for sensitivity in the detection and titration of murine cytomegalovirus antibody. The three tests were compared using sera from experimentally inoculated and naturally infected mice bled at intervals from 3 to 140 days postinfection. In the acute infection, complement fixation and indirect immunofluorescent assay tests were of comparable sensitivity for early detection of antibody, whereas the ELISA was less sensitive. In persistent infection, higher titers were recorded with ELISA. Since murine cytomegalovirus has been shown to exert significant effects on the immune response of infected mice, this antigen should be included routinely in viral antibody screening programs.     

河南部分规模化猪场猪传染性胸膜肺炎的血清学调查

利用间接血凝试验对来自河南省部分地区未接种过传染性胸膜肺炎疫苗的部分规模化猪场的807份血清样品进行了传染性胸膜肺炎的抗体检测，以抗体效价≥1∶8为阳性作为判定标准，结果发现被检样品的总阳性率为30.11%（243/807），经产母猪、40日龄断奶仔猪、90～120日龄猪和160～180日龄猪的阳性率分别为56.94%（123/216）、9.04%(17/188)、27.65%(60/217)、23.12%(43/186)，上述各阶段阳性猪抗体效价的几何平均数分别为1∶65.15、1∶11.53、1∶26.92、1∶19.73。     

猪血凝性脑脊髓炎病毒的血清学调查

应用血凝和血凝抑制试验检测了从辽宁省部分地区采集的猪血清中血凝性脑脊髓炎病毒（HEV）的抗体滴度。结果表明,368份血清样品中,有162份呈抗体阳性反应,阳性率高达44.0%。被采集血清的猪未表现出临床症状,说明该地区存在血凝性脑脊髓炎病毒的隐性感染。     

Enzyme-linked immunosorbent assay for the detection of toxoplasmosis in swine: interpreting assay results and comparing with other serologic tests

An enzyme-linked immunosorbent assay (ELISA) was developed for the serodiagnosis of swine toxoplasmosis. The results of the ELISA were interpreted as percentage (% ELISA) and ELISA titer and were compared with results of the dye test, indirect fluorescent antibody test, and indirect hemagglutination test. The results obtained from both the % ELISA and ELISA titer directly correlated with the dye test results. The ELISA is a rapid, sensitive, and readily performed serologic test that should be ideal for examining (screening) swine for toxoplasmosis.     

Antibody response to modified-live canine adenovirus vaccine in African hunting dogs (Lycaon pictus).

Nine hunting dogs were bled pre and post vaccination with modified-live virus vaccine. Their antibody responses to canine adenovirus type 1 was measured by an indirect immunofluorescent antibody test. The vaccine proved to be both effective and safe.     

巴楚蘑菇多糖对鸡免疫功能的影响

采用细菌吞噬试验、体内淋巴细胞转化试验、血凝和血凝抑制试验来研究巴楚蘑菇多糖对鸡免疫功能的影响.结果显示巴楚蘑菇多糖能够提高鸡白细胞的吞噬能力、淋巴细胞的转化率及抗体效价.结论巴楚蘑菇多糖具有免疫增强作用.