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.     

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.     

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.     

An indirect fluorescent antibody test for the detection of Cytauxzoon-like organisms in experimentally infected cats.

Antiserum to feline Cytauxzoon-like parasites was used in conjunction with labeled rabbit antisera to feline globulins to detect the presence of Cytauxzoon-like parasites in spleens of experimentally infected cats. Frozen spleen sections from 21 infected cats showed positively fluorescing masses within splenic veins and a diffuse scattering of discretely fluorescing cells in the red and white pulp. The distribution of fluorescence corresponded with the appearance of parasitized reticuloendothelial cells in histological preparations of spleen tissue. This indirect fluorescent antibody test consistently detected the presence of Cytauxzoon-like parasites in frozen spleen sections from experimentally infected cats.     

An indirect fluorescent antibody test for the detection of antibody to swine infertility and respiratory syndrome virus in swine sera.

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)     

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.     

Use of the indirect fluorescent antibody test in the detection of bovine respiratory syncytial virus antibodies in bovine serum.

The indirect fluorescent antibody test was adapted for identifying bovine respiratory syncytial virus and its specific antibody, using goat turbinate (GTU) cells. The virus caused maximal cytopathic effects in GTU cells 4 to 8 days postinfection, but fluorescence was not readily detected during this period. Fluorescence was maximal in infected GTU cells at 24 to 36 hours postinfection, but could be detected 48 hours postinfection. Bovine serums (331) which had been submitted to the Oklahoma Animal Disease Diagnostic Laboratory were tested for antibodies to this virus, and 73.6% were found to be positive.     

Prevalence of antibodies to Dermatophilus congolensis in sheep and goats in Nigeria

Summary

A serological survey on dermatophilosis was carried out amongst sheep and goats in Kaduna State of Nigeria. Sera were‐obtained from slaughter animals and from sheep kept on an isolated ranch. The percentage of seropositive animals was 28.0 in slaughter sheep, 0.0 in sheep kept on the ranch, and 23.2 in slaughter goats. The high prevalence of D. congolensis antibodies among small ruminants compares well with the level of prevalence reported of cattle and calls for a concerted government effort for the control of the disease.     

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.     

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.     

Prevalence of antibodies to Dermatophilus congolensis in sheep and goats in Nigeria

A serological survey on dermatophilosis was carried out amongst sheep and goats in Kaduna State of Nigeria. Sera were obtained from slaughter animals and from sheep kept on an isolated ranch. The percentage of seropositive animals was 28.0 in slaughter sheep, 0.0 in sheep kept on the ranch, and 23.2 in slaughter goats. The high prevalence of D. congolensis antibodies among small ruminants compares well with the level of prevalence reported of cattle of cattle and calls for a concerted government effort for the control of the disease.     

A modified indirect immunofluorescent assay for the detection of antibody to Eperythrozoon ovis in sheep

Experimental ovine eperythrozoonosis was studied using Giemsa staining of blood films and a modified indirect immunofluorescent antibody assay (IFAA). The serums of 21 Border Leicester Merino cross lambs between 12 weeks and 7 months-of-age were analysed before and after infection with Eperythrozoon ovis (E. ovis) using the IFAA test. No rise in the IFAA titre was seen until day 7 and this coincided with the first detection of E. ovis organisms in blood smears stained with Giemsa. The percentage of E. ovis infected red blood cells peaked on day 14, but the IFAA titre did not peak until day 35. Titres to E. ovis, on average, had begun to drop by day 63. There was considerable individual variation in response to E. ovis infection as measured by the IFAA. Titres as high as 6,400 were observed in individual sheep at the peak of E. ovis parasitaemia of red cells. One sheep had a titre of 51,200 nineteen days after infection, and titres of 3,000 were maintained for several months in a few sheep. The assay proved reliable, and up to 100 samples per day could be tested. The antigenicity of the slide preparations was found to be satisfactory after storage for 6 months at -20 degrees C and 4 degrees C and for 28 months at -70 degrees C. Temperature fluctuations during storage rendered slides unsuitable for the IFAA after these times. A method of storing E. ovis infected blood in liquid nitrogen is described.     

An indirect flow cytometric test for detection of anti-neutrophil antibodies in dogs

OBJECTIVE: To develop a clinically applicable assay for detection of serum anti-neutrophil antibodies in dogs. SAMPLE POPULATION: Serum samples of 20 healthy dogs and 20 sick dogs. PROCEDURES: An indirect immunofluorescence assay was developed in which canine serum was incubated with paraformaldehyde-fixed neutrophils and subsequently incubated with fluorescein-conjugated rabbit anti-dog IgG. Neutrophil median fluorescence intensity and the percentage of neutrophils with an increase in fluorescence intensity were determined by use of a flow cytometer. RESULTS: Neutrophils incubated with serum from healthy and sick dogs had a normally distributed curve when displayed as a histogram. Alloantibodies or immune complexes that significantly affected test results were not detected. Hyperglobulinemia did not appear to affect test results. The neutrophil donor did not significantly affect test results. With 1 exception, results for the sick dogs did not differ appreciably from those for healthy dogs. Serum from a dog with steroid-responsive neutropenia had a greater neutrophil fluorescence value and percentage of neutrophils with an increase in fluorescence intensity, compared with either healthy or sick dogs. CONCLUSIONS AND CLINICAL RELEVANCE: The indirect immunofluorescence test gave consistent results for healthy and sick dogs and detected anti-neutrophil antibodies in a dog with steroid-responsive neutropenia. Definitive evaluation of the test will be dependent on evaluation of persistently neutropenic dogs and correlation of test results with a response to immunosuppressive therapy.     

An indirect haemagglutination test for the detection of Brucella ovis antibodies

Abstract

Extract

Surveys on perinatal infection in lambs in New Zealand have been reported and the pathology and bacteriology of the conditions described (Hartley and Boyes, 1955 Hartley, W. J. and Boyes, Betty W. 1955. Proc. N.Z. Soc. anim. Prod., 15: 120–120.  [Google Scholar], 1964 Hartley, W. J. and Boyes, Betty W. 1964. N.Z. vet J., 12: 33–33. [Taylor &; Francis Online] , [Google Scholar]; McFarlane, 1955 McFarlane, D. 1955. Proc. N.Z. Soc. anim. Prod., 15: 104–104.  [Google Scholar]; Hartley and Kater, 1964 Hartley, W. J. and Kater, Joan C. 1964. N.Z. vet. J., 12: 49–49. [Taylor &; Francis Online] , [Google Scholar]). Potentially pathogenic organisms were isolated from 58 to 288 lambs from five flocks, Clostridium septicum being isolated from five of these cases (Hartley and Boyes, 1955 Hartley, W. J. and Boyes, Betty W. 1955. Proc. N.Z. Soc. anim. Prod., 15: 120–120.  [Google Scholar]). In another survey, 5.5% of lambs born dead or dying up to 4 weeks of age died from navel infection. Clostridium septicum was isolated from 69% of 48 consecutive cases (Hartley and Boyes, 1964 Hartley, W. J. and Boyes, Betty W. 1964. N.Z. vet J., 12: 33–33. [Taylor &; Francis Online] , [Google Scholar]). McFarlane (1955 McFarlane, D. 1955. Proc. N.Z. Soc. anim. Prod., 15: 104–104.  [Google Scholar]) recorded that 7.3% of perinatal mortality was due to navel infection but no bacteriology was carried out nor was the organism suspected stated. On individual farms, up to 15% of lambs recorded died from navel ill. It should be pointed out that, in this survey, only small numbers of lambs were received from some properties.     

The indirect fluorescent antibody test (IFAT) for the detection of Nosema cuniculi antibodies in the blue fox (Alopex lagopus).

During recent years nosematosis has been a major problem in the breeding of blue fox in the Scandinavian countries, causing heavy losses among growing pups (Nordstoga 1972, Nordstoga et al. 1974). The lack of reliable methods for diagnosing the infection in live foxes has so far made epizootiologic studies of the disease very difficult. However, reports on the IFAT in rabbits with nosematosis (Cox et al. 1972, Chalupsky et al. 1971, 1973, 1974), encouraged the search for a method of detecting Nosema antibodies in fox sera.     

An indirect haemagglutination test for the detection of Vibrio fetus antibodies

Abstract

Extract

Indirect bacterial haemagglutination was first reported by Keogh et al. (1947 Keogh, E. V., North, E. A. and Warburton, M. F. 1947. Nature (Lond.), 160: 63–63.  [Google Scholar]). It depends on the adsorption of bacterial antigens to the surface of red blood cells rendering them agglutinable in the presence of homologous bacterial antibody. A comprehensive review by Neter (1956 Neter, E. 1956. Bact. Rev., 20: 166–166.  [Google Scholar]) summarizes the methods used and results achieved with various bacterial antigens. Biberstein (1955 Biberstein, E. L. 1955. Cornell Vet., 46: 144–144.  [Google Scholar]) used an adaption of Neter's method of antigen preparation in studying the antigenic relationships of Vibrio species. The objects of the present studywere to determine whether an erythrocyte adsorbable antigen could be obtained from Vibrio fetus, to compare its sensitivity with a formalinized bacterial antigen, and to study its application to the detection of antibodies in bovine vaginal mucus. The work will be described in two sections, the first dealing with the preparation and properties of sheep red cells modified with material derived :from V. fetus, and the second with the detection of antibodies in bovine vaginal mucus.