Comparison of enzyme-linked immunosorbent assay, indirect fluorescent antibody test, and direct agglutination test for detecting Toxoplasma gondii antibodies in naturally aborted ovine fetuses

Results obtained in an enzyme-linked immunosorbent assay (ELISA), an indirect fluorescent antibody test (IFA), and a modified direct agglutination test (MAT) for Toxoplasma gondii antibodies from examination of fetal fluids from 377 aborted ovine fetuses were compared. Sixty-seven samples were positive by MAT (titers 1:16 to greater than 1:65,536), 58 were positive by ELISA, and 62 were positive by immunoglobulin G-IFA. The MAT was preferred because it required less time, labor, and special equipment. It was simple to run, could be done on serum from any species without modification, and it was more effective than the IFA for detecting toxoplasma antibodies in severely autolyzed fetuses. No advantage was found in determining immunoglobulin M antibodies in ovine fetal sera.     

应用间接荧光抗体技术检测兔波氏杆菌

以兔波氏杆菌为免疫原,强化免疫家兔,制备兔抗血清为第一抗体;以标准的羊抗兔IgG-FITC荧光抗体为第二抗体,建立了检验兔败血波氏杆菌的间接荧光抗体技术.用火焰、甲醇、丙酮三种方法固定标本,分别经过10 min和20 min两种不同时间固定,然后滴加不同稀释倍数的兔抗血清,滴加羊抗兔IgG-FITC,于荧光显微镜下观察.结果表明甲醇固定10 min和火焰固定,兔抗血清抗体效价为1:80时效果最好.     

An indirect fluorescent antibody test for antibodies to transmissible gastroenteritis of swine.

The indirect fluorescent antibody test was modified to provide a rapid technique for the detection, screening and titration of antibodies to transmissible gastroenteritis of pigs. Large numbers of slides containing transmissible gastroenteritis antigen were prepared by planting mixtures of infected and uninfected swine testicular cells onto multiwelled teflon-coated slides. After overnight incubation, about one-half of the cells in each well were infected which provided contrast to aid in detecting specific fluorescence in the presence of varying degrees of background staining. Following fixation, antigen slides were stored at -20 degrees C until used. The indirect fluorescent antibody test was compared to the virus neutralization test in both the screening and titration of swine sera containing transmissible gastroenteritis antibodies. The test was found to be sensitive and reliable and to offer certain advantages over the virus neutralization test.     

The indirect fluorescent antibody test for bovine anaplasmosis

Summary

An indirect fluorescent antibody test was used succesfully for the serodiagnosis of experimental Anaplasma infections in cattle. Specific antibodies were detected three to ten days after anaplasma bodies werd found in the blood, and persistedat least 15 weeks post‐infection.

An American and an African stock of A. marginale were used to prepare antigens, and gave comparable results when tested on sera positive to either of these stocks, as well as to an A. centrale‐like stock from Korea.

There were no cross‐reactions with several Theileria, Babesia, Trypanosoma and Eperythrozoon species.     

Encephalitozoonosis in the blue fox. Comparison between the india-ink immuno-reaction and the indirect fluorescent antibody test in detecting Encephalitozoon cuniculi antibodies

Sera from 32 foxes sampled at intervals varying from 20 to 70 days after oral inoculation with E. cuniculi spores were tested by the india-ink immunoreaction (IIR) and the indirect fluorescent antibody test (IFAT). Using the IFAT, antibodies were detected at low levels in sera sampled on days 20 and 29 post inoculation, whereas the IIR failed to reveal antibodies in the same sera. In sera sampled from day 35 until day 70 post inoculation, antibodies were detected by both tests, the IIR-titres reaching the magnitude of the IFAT-titres after about 50 days posit inoculation.In 14 sera sampled from foxes of at least 46 days of age and with signs of encephalitozoonosis, the tests gave almost identical results.Comparing IIR- and IFAT-determined antibody titres using E. cuniculi antigens of blue fox and rabbit origin in the test, the antigens seemed to be closely related, supporting the suggestion that the isolates are strains of the same microsporidian species.     

The fluorescent antibody and indirect fluorescent antibody assays for diagnosing stunting syndrome of turkeys

The fluorescent antibody (FA) assay was developed for detecting the stunting syndrome agent (SSA) from intestinal tissue. Similarly, the indirect fluorescent antibody (IFA) assay was developed for detecting serum antibodies to SSA. Convalescent antiserum from turkeys orally immunized with SSA was found to be the primary antibody of choice for the FA assay. Intestinal jejunal samples from poults inoculated 3-4 days postinoculation (DPI) was found to be the best antigen source for the IFA assay. SSA was detected from the intestinal tracts of experimentally inoculated birds at 2 DPI with highest level of reactivity at 3 DPI by the FA assay. After 4 DPI the level of SSA infectivity of the intestines waned to low levels. Serum antibody was detected from experimentally inoculated birds as early as 7 DPI with all birds tested seroconverting by 12 DPI. These assays should prove useful for future studies concerning stunting syndrome.     

Influence of medication on development of serum antibody to swine dysentery as detected with indirect fluorescent antibody method.

Serums from 119 swine exposed to swine dysentery inoculum, and medicated with various drugs, were tested for antibodies to the large spirochete, using the indirect fluorescent antibody test, and were compared in tests with known positive serums from 18 nonmedicated swine which had recovered naturally. An inverse relationship existed between the efficacy of the drug and the serum antibody titer (highest dilution of the serum producing immunofluorescence of large spirochetes). The more efficacious drugs or doses resulted in lesser development of serum antibody. Diarrhea usually seemed necessary for the development of serum antibody. With the less efficacious drugs, there were more days of diarrhea. Some swine had diarrhea but did not develop an antibody titer, and a few swine had a titer but did not develop diarrhea. Swine which developed a titer were more immune against reexposure with infective inoculum. The medicaments, especially those given at higher concentrations, seemed to resolve the diarrhea or prevent the development of diarrhea, occurrences which were necessary for the development of immunity.     

Cryptosporidium antibodies in manitoba cattle: a pilot study using an indirect fluorescent antibody procedure

An indirect fluorescent antibody test, using feces-derived oocysts as antigen, was used to detect antibodies to Cryptosporidium spp. in bovine sera in Manitoba. Antibodies were detected in 29 of 50 (58%) sera collected from animals of various ages on farms where calves had laboratory-confirmed cryptosporidiosis and in 76 of 186 (40.9%) sera collected at random from culled breeding stock. Serum antibody, presumably colostral in origin, did not appear to protect young calves from the infection. No geographic preference for the infection was demonstrated.     

Kinetics of antibody response to Ehrlichia canis assayed by the indirect fluorescent antibody method.

The kinetics of antibody production response to experimentally induced infection of dogs with Ehrlichia canis was determined by ion-exchange and molecularsieve chromatography and by indirect fluorescent antibody (IFA) test. The first IFA antibody at 7 days after inoculation resided in immunoglobulin M (IgM) and immunoglobulin A (IgA) classes. At approximately 21 days after inoculation, the antibody was in IgM, IgA, and immunoglobulin G (IgG) classes. Thereafter, antibody concentrations continued to increase in the IgG class; those in the other 2 immunoglobulin classes had a variable pattern. In 2 dogs which died 60 and 114 days after inoculation, a decrease of antibody concentration in the 3 immunoglobulin classes was evident at the time of death. In the carrier dog, however, which was killed 147 days after inoculation, antibody concentrations sustained increasing titers in the 3 immunoglobulin classes.     

Prevalence of antibodies to porcine adenovirus in swine by indirect fluorescent antibody test

An indirect fluorescent antibody test was developed for routine identification of a porcine adenovirus and its specific antibody. Two specific-pathogen-free young pigs were inoculated with the viral antigen prepared in continuous porcine kidney cell cultures, and their sera were used as an antibody reagent to standardize the test. Sera of adult pigs with respiratory problems, obtained from pig farms in Quebec, were tested for antibodies to this virus; 83 of 540 sera tested (15.2%) were found to be positive.     

牛传染性鼻气管炎间接ELISA诊断方法的建立

以牛肾细胞系（MDBK）培养牛传染性鼻气管炎病毒（IBRV）Bartha Nu/67株,经超速离心纯化病毒,再经超声破碎处理后作为诊断抗原,建立了检测牛血清IBRV抗体的间接酶联免疫吸附试验.该ELISA的判定标准为：血清D490 nm值大于0.369的判为阳性,小于0.295的判为阴性,在0.295与0.369之间的为可疑.特异性和重复性试验结果表明,该方法特异性高、重复性好.与法国进口ELISA抗体诊断试剂盒比较,其符合率为96.3%;与中和试验比较,符合率为95.8%,且敏感性更高.应用该诊断方法调查了我国部分地区IBRV的感染情况,结果显示,这些地区的IBRV感染率为67.1%.     

Brucella abortus-associated meningitis in aborted bovine fetuses.

Granulomatous meningitis was present in 6/33 bovine fetuses from which Brucella abortus (B. abortus) had been isolated. Meningitis was severe in three fetuses, moderate in one fetus, and mild in the remaining two fetuses. The meningitis was characterized by the infiltration of a mixed population of lymphocytes, plasma cells, and macrophages in the leptomeninges. Vasculitis characterized by the infiltration of lymphocytes and plasma cells in the vascular wall was observed in the vessels of the cerebral cortices of 4/6 fetuses. Gram negative coccobacilli were present in the cytoplasm of the leptomeningeal macrophages and extracellularly. Brucellar antigens labeled by the avidin-biotin-peroxidase complex method were present in massive amounts in leptomeningeal macrophages and in small foci of stained cells in the choroid plexus and ependyma. The findings indicate that B. abortus is one of pathogens capable of inducing meningitis in bovine fetuses.     

Identification of Babesia bigemina in the tick Boophilus decoloratus by the indirect fluorescent antibody technique.

Developing stages of Babesia bigemina were detected in the Giemsa-stained haemolymph smears of replete Boophilus decoloratus females engorged on infected animals. Replicate smears of these were prepared for staining by the indirect fluorescent antibody (IFA) technique. With specific antisera to B bigemina in dilutions up to 1/160 and rabbit antibovine globulin conjugated with fluorescein isothiocyanate (conjugate) the B bigemina stages were seen to fluoresce under the fluorescent microscope. When antisera against cattle Theileria spp or negative control sera were used, fluorescence was not detected in dilution above 1/5 and there was a complete absence of fluorescence when the conjugate alone was used. Thus the developing stages of B bigemina from the haemolymph could be identified using the IFA technique. Both spherical and elongated developing stages were seen to fluoresce specifically. The apical and the perinuclear regions and the posterior end of the vermicules appeared to fluoresce more intensely than the rest of the cytoplasm.