共查询到20条相似文献，搜索用时 31 毫秒
1.
K J Reis 《Veterinary immunology and immunopathology》1989,23(1-2):129-137
A hemolytic assay was developed for the measurement of functional equine complement activity. The assay utilizes antibody sensitized chicken erythrocytes as the target cell and was specific for classical pathway (antibody dependent) complement activity. The assay was found to be reproducible and more sensitive than previous reports using other species of target cells. Total serum complement (CH50) values were determined for five mares and their foals and followed over a period of 3 months. 相似文献
2.
Concentrations of bovine factor B (Bbov) were determined by radial immunodiffusion in sera of 46 Holstein cows and heifers aged one to nine years. Mean values were 34.2 +/- 5.3 mg/100 ml. A hemolytic diffusion plate assay in agarose gel in presence of 10 mM EGTA and 5 mM Mg accurately measured concentrations of purified Bbov but gave higher mean values, i.e. 47.8 +/- 10.2 mg/100 ml, for concentrations of Bbov in whole sera. Hemolytic values obtained by the hemolytic diffusion plate assay, however, weakly correlated (r = .4539, p less than 0.01) with the serum concentration of Bbov measured by radial immunodiffusion. It was concluded that the hemolytic diffusion plate assay was not an accurate technique for the quantitative measurement of Bbov but a good assay for quantitation of the total hemolytic activity mediated via activation of the alternative complement pathway. It is suggested that the difference between the values obtained by the two tests for one particular serum is, to some degree, an expression of the ratio of amplification and restriction of the alternative pathway activity. No significant heritability (offspring and one parent) was detected for the hemolytic activity of serum. A heritability of 0.93 at a significance level of p less than 0.1 was determined for the serum concentration of Bbov. 相似文献
3.
An indirect hemagglutination test was developed for the diagnosis of canine schistosomiasis. Specific-pathogen-free dogs and dogs infected with a variety of other helminths were experimentally infected with Heterobilharzia americana. Sera collected at various times after the dogs were exposed to cercariae were evaluated. Glutaraldehyde-fixed sheep erythrocytes sensitized with whole adult worm antigens provided a stable antigen substrate when tested over a period of months. The test produced sensitive and specific diagnosis at an appropriate serum dilution. Serum dilutions less than 1:80 from dogs with other helminth infections resulted in nonspecific agglutination. By postexposure day 60, coinciding with the end of the prepatent period, serum titers of dogs infected only with H americana equaled, and then surpassed, those of the other dogs. 相似文献
4.
A 51Cr release microhemolytic complement assay is described to detect hemolytic complement activity in bovine milk. 51Cr-labeled guinea-pig erythrocytes (GPRBC), which have been sensitized with a subagglutinating amount of rabbit anti-GPRBC, are placed in microtiter plates. Pooled bovine sera as source of complement to achieve about 50% of 51Cr release were added to each well prior to the addition of the samples on the test. Determination of CH100 titer was obtained by difference of counting between heated and unheated diluted whey samples from a standard linear regression. Comparative hemolytic values throughout lactation were established for the first time and confirmed the improved sensitivity of the assay. 相似文献
5.
The use of a passive hemolysis system to evaluate the complement activities of six mammalian species
A passive hemolysis assay system was developed which permitted comparisons of the hemolytic activities of complement (C) from six species. This system employs a single antigen and an antiserum raised in one species. Thus, variations resulting from different target antigens and those inherent in using antibodies (of different affinities and isotypes) raised in a variety of species were minimized. Of the erythrocytes (E) examined, those from horses and guinea pigs were most susceptible to lysis, and either would be suitable, as a tentative choice, for measuring C activity of a previously unstudied species. Horse serum had the lowest C activity of any of the sera tested. It lysed certain cells only at high concentrations, and the hemolytic activity dropped off sharply with minimal dilution. The data presented in this paper could also be used for selecting target E for C studies using direct lysis with antibodies raised against the cells. 相似文献
6.
《Comparative immunology, microbiology and infectious diseases》2014,37(5-6):305-311
Arthrographis kalrae is a dimorphic, cosmopolitan and neurotropic fungus that has been described as a rare human pathogen. This study investigated the hemolytic and cytotoxic activities of A. kalrae cell-free antigens (CFAs). Total CFAs and their Sephadex chromatography fractions were tested on mouse erythrocytes for hemolysis and on the P3U1 cell line for cytotoxicity. Hemolytic and cytotoxic activities were detected in distinct molecular mass (MM) fractions. Additionally, antibodies against isogenic erythrocytes sensitized with CFAs (anti-E-CFAs) inhibited hemolysis but not cytotoxicity. Hemolysis was not affected by heating, and a higher reactivity was detected in the carbohydrate-rich fractions, which decreased after reduction by periodate treatment. The pioneering nature of this work is due to the demonstration of the cytotoxic activity in A. kalrae and the suggestion that this activity may be due to molecules distinct from the hemolytic factor, with the latter potentially being a component with a high MM. 相似文献
7.
Bovine serum and colostral whey samples were examined for lysozyme and haemolytic complement activity, employing agar plate techniques. The tests were carried out in agarose gel containing Micrococcus lysodeikticus (for lysozyme), and antibody sensitized rabbit erythrocytes (for complement), respectively. The confirmation of lysozyme (E.C.3.2.1.17) — dependent lysis has been presented elsewhere (Lie & Syed 1986), while heat-inaotivation and antibody to C3 were used in the present study to confirm that the haemolytic activity was attributable to the complement cascade. Repeatability and sensitivity of the described tests were found to be superior to those of photometric procedures. Staining and preservation techniques were developed which extended the applicability of the assay, as they made reading of results independent of time and resulted in the plates being very suitable for photography and storage. 相似文献
8.
A serologic method for the detection of Corynebacterium pseudotuberculosis infections in horses 总被引：2，自引：0，他引：2
H D Knight 《The Cornell veterinarian》1978,68(2):220-237
A serologic technique useful for detecting antibodies formed in horses in response to infection with Corynebacterium pseudotuberculosis is described. The test relies on the ability of C. pseudotuberculosis toxin to produce a wide zone of hemolysis when applied to erythrocytes previously treated with a sterile filtrate of Corynebacterium equi broth culture. The synergistic hemolytic activity can be neutralized by anti-C. pseudotuberculosis serum. This test was used to analyze sera from 616 horses for the presence of C. pseudotuberculosis antitoxin. Of 177 animals (see Table 2) found positive, there were 34 horses with bacteriologically confirmed, active infections and 18 with active but unconfirmed infections. In addition, 13 animals had a history of having had the disease and 112 had no history or evidence of having had the infection. The other 439 horses had negative titers. Statistical treatments confirmed the value of the test as an epidemiological tool but precluded using only titers for the diagnosis of active clinical disease. 相似文献
9.
A simple, one-step, alternative pathway (AP) hemolytic assay for bovine C3 has been developed. Methylamine was used to prepare a bovine serum reagent, R3, functionally depleted of C3. The addition of purified bovine C3 to the R3 reconstituted, in a dose-dependent manner, the hemolytic activity for unsensitized heterologous erythrocytes. The assay was used to determine relative levels of C3 in different bovine serum samples. Human C3 and bovine C3 were interchangeable in the assay. Reconstitution of bovine and human R3 reagents with homologous or heterologous C3, in the presence of different species of erythrocytes, provided evidence that cell surface regulation of the homologous hemolytic AP may not be limited to the assembly and activity of the C3 convertase. The AP assay was more sensitive and less complex to perform than a standard classical pathway assay for bovine C3. 相似文献
10.
Optimal conditions for assaying hemolytic complement of goat (caprine) and swine (porcine) sera were determined. Effects of the following were tested: pH, ionic strength, calcium and magnesium ion concentrations, time and temperature of incubation, and ethylenediamine tetracetate concentration. Guinea pig erythrocytes sensitized with goat or cattle antibodies were the most sensitive target cells for goat complement. Sheep and cattle erythrocytes sensitized with rabbit hemolysin were the best target cells for swine complement. Barbital buffer, pH 7.3, ionic strength of 90 nmM relative salt concentration, containing 0.5 mM CaCl2 and 1 mM MgCl2 was the best for swine complement assay. Goat complement lysed best in a barbital buffer, pH 8, ionic strength of 90 to 120 mM of relative salt concentration, in presence of 0.5 mM CaCl2 and 1 mM MgCl2. The optimal incubation temperature was 37 degrees C for both complements. The complement dependent lysis required 75 minutes to reach its maximum. Ethylenediamine tetracetate in 4 mM concentration completely inhibited lysis by both species complements. The CH50 for goat sera varied between 18 and 75 per ml, in swine sera between 75 and 210 per ml. 相似文献
11.
L.R. Ellingsworth C.A. Holmberg B.I. Osburn 《Veterinary immunology and immunopathology》1983,5(2):141-149
A microtiter system was used to measure hemolytic complement levels in serum from eleven nonhuman primate species. The species studied were (rhesus macaque), (bonnet macaque), (pig-tailed macaque), (crab-eating macaque), (stumptailed macaque), (yellow baboon), (olive baboon), (African green monkey), (owl monkey), (spider monkey), and (thick-tailed galago).The optimal hemolytic complement titer of the various nonhuman primate species was found to vary with different species sources of erythrocytes and anti-erythrocyte reagents used in the assay. No single erythrocyte and anti-erythrocyte test reagent produced optimal titers for all of the primate species examined. Sera from several species was found to have high spontaneous lytic activity towards non-sensitized sheep erythrocytes which for six species ( and ) was equal to the titer for antibody sensitized erythrocytes. Evidence of alternate pathway complement activation as a possible reason for the high titer of lytic activity towards unsensitized erythrocytes could not be demonstrated for any nonhuman primate species. In one species, , the sensitizing activity of normal serum for sheep erythrocytes was shown to be in the IgM containing fraction obtained with gel filtration and to be absorbed by boiled sheep erythrocyte stroma which contains Forssman antigen. 相似文献
12.
试验旨在利用胶体金免疫层析技术建立快速检测犬血清中犬细小病毒（canine parvo virus, CPV）血凝抑制（haemagglutination inhibition, HI）抗体效价的方法,用于CPV疫苗免疫效果评价。采用双抗体夹心法,以抗CPV血凝相关抗原的单克隆抗体制备CPV抗原检测试纸条;将犬血清进行不同比例系列稀释后,分别与定量CPV抗原充分反应,滴入CPV胶体金试纸条,根据试纸条检测线（test line,T线）消失时的血清最高稀释倍数判断血清中CPV抗体的HI效价;用此方法检测86份犬血清样品,并与传统血凝抑制试验方法进行分析比较。结果显示,成功制备CPV抗原检测试纸条,确定了试纸条检测犬血清CPV-HI效价的反应条件和结果判定标准。结果表明,在检测不同稀释倍数犬血清反应后的CPV抗原时,能使试纸条T线消失时的血清最高稀释倍数与HI效价具有正相关性,犬血清最高稀释倍数乘以4即为HI效价;两种方法的符合率达90.7%。本试验初步建立了胶体金试纸条检测CPV血凝抑制效价的方法,为检测CPV-HI效价提供了一种操作简单、快速的试验方法,可用于CPV疫苗免疫效果评价。 相似文献
13.
MA Yong-ying SUN Ming SHEN TU Fen-qin KONG Han-jin YANG Xin-yan QIN Ya-man CHEN Xi-zhao JIA Hong 《中国畜牧兽医》2016,43(8):1983-1988
The study was aimed to use colloidal gold immune chromatography technology to establish a rapid method for detection of canine serum canine parvovirus (CPV) hemagglutination inhibition (HI) titer and CPV vaccine immunization effect assessment.Double antibody sandwich method and monoclonal antibodies of anti-CPV hemagglutination antigen were used to prepare CPV antigen test strip.Canine serum with different proportion respectively was mixed with quantitative CPV antigen for full reaction,then dropped the mixture into the CPV colloidal gold test strip,so according to the highest serum dilution ratios when the test strip line T (line T) vanishes,it was to judge CPV antibodies in serum of the HI titer.This method had been used to detect 86 canine serum samples,at the same time,analyzing and comparing it with traditional hemagglutination inhibition test method.The results showed that the CPV antigen detection test strip was successfully prepared,and the reaction conditions and results of the test strip for detecting the titer of CPV-HI in canine serum were determined.The results indicated that when detecting CPV antigen after the dilution of different ratios of canine serum,the highest serum dilution ratios when the strip line T vanished and the HI titer had positive correlation.The highest dilution ratios of canine serum multiplied by 4 was the HI titer.The results of two methods had 90.7% consistency.This experiment established the colloidal gold immune chromatography test strip for the detection of CPV-HI titers method initially.This CPV-HI detection provided a simple and fast test method for the effect evaluation of CPV vaccine immune. 相似文献
14.
Antisera to sheep erythrocytes (E) were raised in cattle, rabbits, mice, hamsters, guinea-pigs, ferrets, badgers, hedgehogs and fowls. Cross activation of total haemolytic complement (THCA) examined all combinations of sensitized sheep E and normal sera (including human); kinetic assays examined the lysis of E sensitized with rabbit antibodies. From the same species, all combinations of normal serum and xenogeneic E were used to measure total alternative pathway activity (TAPA); TAPA was also activated by rabbit and sheep E in titrations and in agarose gels, and examined kinetically against rabbit E. Ox, rabbit and fowl sera were low in THCA, guinea-pig complement was universally active, while human complement showed marked selectivity; ferret, badger and hedgehog sera were activated to high titres but probably via the alternative pathway. In studies of TAPA an inverse relationship existed between serum complement activities and the activating abilities of E from the same species. The most efficient activators of alternative pathway were E from rabbits and laboratory rodents, while the sera with broadest response were badger, ferret and fowl. Kinetic studies of TAPA showed that initiation of lysis and subsequent completion of lysis could occur with different efficiencies, suggesting these events reflected separate events in complement activation. 相似文献
15.
Evaluation of heat-sensitive, neutrophil-toxic, and hemolytic activity of Haemophilus (Actinobacillus) pleuropneumoniae 总被引：28，自引：0，他引：28
S Rosendal J Devenish J I MacInnes J H Lumsden S Watson H Xun 《American journal of veterinary research》1988,49(7):1053-1058
Cytotoxic and hemolytic activity of Haemophilus (Actinobacillus) pleuropneumoniae serotype 1 strain CM5 was investigated because of the potential role as a virulence determinant. Viable bacteria were toxic for porcine and bovine neutrophils, whereas bacteria killed by heat treatment at 60 C for 1 hour were not. Similarly, bacteria-free culture supernatant was cytotoxic and hemolytic in assays that used porcine neutrophils and erythrocytes, whereas supernatant treated at 60 C for 1 hour had no activity. Erythrocytes from various species were susceptible to the hemolytic activity of bacteria-free culture supernatant, with ovine and bovine erythrocytes being most sensitive. The neutrophil-toxic and hemolytic activity of bacteria-free culture supernatant was inhibited by cholesterol and oxygen and abolished after trypsin digestion. The neutrophil-toxic and hemolytic activity was preserved during storage at or less than 4 C, but was lost rapidly at 56 C or 80 C. Neutralizing antibodies were demonstrated in serum of pigs and rabbits immunized with 10-fold concentrated culture supernatant of strain CM5 and in field pigs that had recovered from natural infection with H pleuropneumoniae serotype 1. Bacteria-free culture supernatants of 18 strains, including H pleuropneumoniae serotypes 1 through 10, Actinobacillus suis, and Haemophilus taxon minor group, were tested for heat-sensitive, neutrophil-toxic, and hemolytic activity. Fifteen strains were neutrophil toxic, but only 10 of these were hemolytic. Haemophilus pleuropneumoniae, serotype 1, strain VLS557; serotype 5, strain K17; and Haemophilus taxon minor group strain 33PN were neither cytotoxic nor hemolytic. 相似文献
16.
Recombinant LipL32 antigen-based single serum dilution ELISA for detection of canine leptospirosis 总被引：4，自引：0，他引：4
Dey S Mohan CM Kumar TM Ramadass P Nainar AM Nachimuthu K 《Veterinary microbiology》2004,103(1-2):99-106
A recombinant antigen-based single serum dilution enzyme-linked immunosorbent assay (ELISA) was developed to measure the specific antibody activity in sera of dogs with leptospirosis. The recombinant antigen developed and used in the assay was specific for the pathogenic serovars of Leptospira. A linear relationship was found to exist between the predicted antibody titres at a single working dilution of 1:1000 and the corresponding observed serum titres as determined by the standard serial-dilution method. Regression analysis was used to determine a standard curve from which an equation can be derived that allows demonstration of the mentioned correlation. The equation was then used to convert the corrected absorbance readings of the single working dilution directly into the predicted ELISA antibody titres. The assay was proved to be sensitive, specific and accurate as compared to the standard microscopic agglutination test (MAT). 相似文献
17.
Mary Beth Callan Leonard T. Jones Urs Giger 《Journal of veterinary internal medicine / American College of Veterinary Internal Medicine》1995,9(4):277-279
Alloantibodies to high-frequency red cell antigens, defined as inherited traits occurring in 92% to 99% or more of the general population, are recognized as a cause of hemolytic transfusion reactions in humans. Here we describe a dog (dog erythrocyte antigen [DEA] 1.2-and DEA 4-positive) sensitized by prior blood transfusion, for which a compatible blood donor could not be found; transfusion of DEA 1.1-negative blood resulted in hemolytic transfusion reactions. Patient serum from days 1 (before first transfusion) and 16 was available for further testing; using 4 dogs with different blood types as potential donors, the major crossmatches were compatible using serum from day 1. However the crossmatches were all incompatible with serum from day 16, indicating that the patient was sensitized to an antigen after the first transfusion. The presence of an alloantibody against DEA 1.1 was not ruled out in this patient, but the incompatibility reactions of patient serum with red cells from donors negative for DEA 1.1 indicated that an alloantibody against a red cell antigen other than DEA 1.1 or any other known DEA for which typing reagents were available (DEA 3, 5, and 7) was present. Subsequently, red cells from 1 of the patient's siblings (DEA 1.2-, 4-, and 7-positive) were found not to agglutinate when incubated with patient's serum from day 16, ruling out the presence of an anti-DEA 7 antibody, and suggesting that an alloantibody against a common red cell antigen missing in the patient and sibling was responsible for the blood incompatibility reactions. Failure to obtain a compatible crossmatch with several universal donors in a dog previously transfused should raise a suspicion that an alloantibody to a common red cell antigen may exist and that a sibling may be a source of compatible blood. 相似文献
18.
OBJECTIVE: To assess whether dogs with experimentally induced type I hypersensitivity against soy protein would respond to soy hydrolysate and develop cutaneous or gastrointestinal tract reactions after intradermal and oral challenge exposure. ANIMALS: 12 na?ve Beagle pups (9 sensitized and 3 control dogs). PROCEDURE: 9 dogs were sensitized against soy protein by administration of allergens during a 90-day period. After the sensitization period, serum concentrations of soy-specific IgE were determined and an intradermal test was performed to confirm the dogs were sensitized against soy protein. An intradermal challenge test and an oral challenge test with native and hydrolyzed soy protein were conducted on 6 sensitized and 2 control dogs. RESULTS: High serum concentrations of soy-specific IgE and positive results for the intradermal test were observed for the 9 sensitized dogs after completion of the sesitization process. Sensitized dogs challenge exposed with hydrolyzed soy protein had a reduced inflammatory response after intradermal injection and no clinical response after an oral challenge exposure, compared with responses after intradermal and oral challenge exposure with native soy protein. CONCLUSIONS AND CLINICAL RELEVANCE: Soy-sensitized dogs did not respond to oral administration of hydrolyzed soy protein. Thus, hydrolyzed soy protein may be useful in diets formulated for the management of dogs with adverse reactions to food. 相似文献
19.
The in vitro hemolytic activity of Tritrichomonas foetus was investigated. The parasite was tested against human erythrocytes of groups A, B, AB, and O, and against erythrocytes of nine adult animals of different species (the rabbit, rat, chicken, cat, dog, swine, horse, bovine, and sheep). The results showed that T. foetus strains (ATCC KV1, K, PAL, 5022, RJ, 90) did not present any hemolytic activity against any human erythrocyte group nor against rabbit, rat, chicken, cat, dog and swine erythrocytes. T. foetus strains, however, lysed horse, bovine, and sheep erythrocytes. No hemolysin released by the parasites could be identified. Hemolysis did not occur with trichomonad culture supernatants, with sonicated extracts of T. foetus, nor with killed organisms. Scanning electron microscopy (SEM) showed that human erythrocytes did not adhere to the trophozoites, in contrast horse erythrocytes adhered to the surface of the parasites and were phagocytosed for up to 90 min. The parasites are able to exert their cytopathic effects through: (a) physical contact established between the two cell surfaces, (b) toxins released from parasites into the interaction media, or (c) the association of both mechanisms. Further studies are necessary to clarify the importance of the hemolytic activity in the biology of T. foetus. 相似文献
20.
Human erythrocytes sensitized with an agglutinating dose of either sheep anti-human erythrocyte antiserum, IgM, slow or fast γ-globulin antibody are haemolysed by foetal lamb or sheep serum complement (C) in the presence of ethyleneglycoltetraacetic acid (EGTA) or Mg2+-EGTA but not ethylenediaminetetraacetic acid (EDTA) or triethylenetetraminehexaacetic acid (TTHA). Dialysis of the C source against phosphate buffered saline rendered it inactive, but addition of Mg2+ or Mg2+-EGTA restored haemolytic activity. Haemolysis of sensitized erythrocytes took place more rapidly in buffer containing Ca2+ and Mg2+ than in EGTA or Mg2+-EGTA. Heating the C source at 50 °C for 30 min or 56 °C for 3 min abolished haemolytic activity. In the presence of EGTA or Mg2+-EGTA, haemolytic activity was restored to 50 °C-heated serum by a CM-cellulose fraction that contained sheep factor B.These results show that sheep C, in the presence of EGTA or Mg2+-EGTA will haemolyse human erythrocytes heavily sensitized with sheep amboceptor. The alternative pathway of sheep C is thought to be involved. 相似文献