• Korean Journal of Veterinary Research
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• v.30 no.3
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• pp.297-307
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• 1990

To establish an agar-gel immunodiffusion (AGID) test for detection of antibodies to Aujeszky's disease virus(ADV) in swine, the precipitating antigens were prepared by four procedures using the Aujeszky's disease virus, NYJ-1-87 strain isolated from the affected piglets in Korea. The optimal condition for AGID test and the properties of the antigens were investigated. To determine the optimal concentration of antigens, four antigens were experimentally prepared by concentrating the viral fluids by 1/30 to 1/200. It was proved that the antigen precipitated with ammonium sulfate at concentration of 1/100 was the most efficient to detect ADV antibodies by AGID test. When the relationship between the concentration of the antigens and the size of precipitating in radial immunodiffusion test was investigated, a high correlation coefficiency at r=0.95 (y=0.23x+23.4) was estimated, In study on the effects of various buffered salt solutions and agars on the sensitivity of AGID test by using the experimental ADV antigens, it was found that 0.05M tris buffer without sodium chloride at pH 7.2 induced the most distinctive precipitating lines, and that there was no significant differences in the sensitivity between the agarose and Noble's special agar. When the efficiency of AGID test was compared with serum neutralization(SN) test, the sensitivity of AGID test was 100% in SN titer over 1 : 16, 91.7% in SN titer of 1 : 8 and 57.1% in SN titer of 1 : 4. The specificity of AGID test compared with the sera with SN titer under 1 : 2 was 98.4%. Protein analysis of the antigens by SDS-PAGE indicated that antigen I and antigen III showed a specific band of polypeptides with molecular weight of 116 K in comparison with the control antigen. Antigen IV, treated with tween-80 and ammonium sulfate, revealed specific polypeptides bands at the molecular weights 45K, 98K and 150 K.

• The Korean Journal of Zoology
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• v.23 no.1
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• pp.1-12
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• 1980

Changes and possible origin of haemolymph proteins during the cuticle formation and hardening are determined by means of acrylamide gel electrophoresis and immunodiffusion. The results by acrylamide gel electrophoresis showed at least 19 protein bands in the haemolymph and 13 fractions in the fat body with relatively constant pattern during the period of cuticle formation and hardening. Both haemolymph and fat body proteins are generally characterized by the presence of three to four heavy stained bands and several thin bands near the top region of the gel. At least over five haemolymph proteins are constantly present during this period. Immunodiffusion tests show that of total eight to nine pupal haemolymph proteins two proteins were already detected in the fat body before pupation and other two proteins were also found in the fat body immediately after pupation, suggesting fat body as possible source of these two haemolymph proteins.

• YAKHAK HOEJI
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• v.41 no.3
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• pp.352-358
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• 1997

The study was carried out on the biochemical characters of Cd-BP(I) after isolation and purification of the protein from the liver of rat injected intraperitoneally with Cd. A c ontinued study has been doing whether Cd-BP(I) could be induced by Cd or by other metals such as Zn and Cu. Antisera were made against the Cd-BP(I) from NewZealand white rabbits. Carried out were ${\gamma}$-globulin purification, then Ouchterlony test adn gel immunodiffusion test. Cd-BP(I) was also found in normal tissues of rat. It was induced up to a considerable level by Cd, whose induced level was higher than that of Cu or Zn treatment. The level of induction by Cu or Zn pretreatment plus Cd treatment was lower than that by simple treatment of Cu or Zn. Such a result was presumably related to the Cd toxicity.

• YAKHAK HOEJI
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• v.43 no.5
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• pp.591-597
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• 1999

The study was carried out on the biochemical characters of Cd-BP(II) after isolation and purification of the protein from the liver of rat ip injection with Cd. A continued study has been doing whether Cd-BP(II) could be induced by Cd or by the other metals such as Zn and Cu. Antisera were made against the antigen of Cd-BP(II) from New Zealand white rabbits. We carried out g-globulin purification, then Ouchterlony test and gel immunodiffusion test. Cd-BP(II) was also found in normal tissues of rat. It was induced up to a considerable level by Cd, whose induced level was higher than that Cu or Zn treatment. The level of induction by Cu or Zn pretreatment plus Cd treatment was lower than that by single treatment of Cu or Zn. Such a result was presumably related to the Cd toxicity.

• Korean Journal of Food Science and Technology
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• v.20 no.6
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• pp.856-861
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• 1988

A new method developed for simultaneous purification of enterotoxin A and C from Staphylococcus aureus strain L 350/1 consisted of chromatography on carboxymethyl (CM)-cellulose using a buffer of variable pH, gel filtration on Ultro gel, and fast protein liquid chromatography(FPLC) using a buffer of variable pH. The enterotoxin A and C were purified by three steps: batchwise adsorption from culture supernatant on Amberlite CG-50; chromatography on CM-cellulose using a buffer of constant pH and molarity; and gel filtration on Sephadex G-75. The purified enterotoxin appeared homogeneous by gel diffusion and polyacrylamide gel electrophoresis. Upon treatment with CM-cellulose using a elution of variable pH, enterotoxin A and C were so close that they were not separated completely. After elution from gels, the enterotoxins appeared as a single peak at the same position. Gel filtration gave a reaction of complete identity to enterotoxin A and C in Ouchterlony immunodiffusion. In FPLC using a CM-cellulose, enterotoxin A and C were simultaneously separated at pH 8.6 and 6.8. When each fraction was performed to gel immunodiffusion, at peak of enterotoxin A and C were not detected each other. In a method of elution by pH-gradient was to be more efficient as a simultaneous separation method in terms of speed, yields and simplicity. The purified toxin A and C were identical to type A and C reference enterotoxin on both disc electrophoresis and Ouchterlony gel diffusion.

• Journal of Veterinary Clinics
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• v.27 no.4
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• pp.402-406
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• 2010

Bovine leukemia virus (BLV) is a delta-retrovirus which causes chronic lymphocytosis in cattle. BLV infections have been divided into two groups such as enzootic bovine leukosis (EBL) and sporadic bovine leukosis (SBL) according to the clinical symptoms in infected cattle. The conventional detection method of BLV was hematological procedure which is determining lymphocytosis in the suspected animals. Recently several sensitive methods were developed to detect antibody to BLV and nucleic acid of the BLV from infected cattle. In this study we have compared the difference of positive rates between agar gel immunodiffusion (AGID) and enzyme linked immunosorbent assay (ELISA) which are using for BLV antibody detection methods. The positive detection rate of ELISA test was 7.4% greater than the positive rate of AGID. The discrepancy of the positive rate between ELISA and AGID were showed in the group of age over one year old to under three year old group. The result from each test agreed very well in the group of over 5 year old cattles. The serological test is very useful method to select the infected cattle for the eradication or control of the disease in the infected herd. But it has a limit by interference of the maternal antibody from the cow of under 6 month old. This study shows that 16.2% of these ages group showed BLV gene positive by polymerase chain reaction (PCR) method. The result suggests that ELISA test need to be used with PCR to clarify misinterpretation of positive animals by antibody response due to the natural infection from maternally derived antibody in calves of under 6 months old.

• Korean Journal of Veterinary Research
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• v.29 no.2
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• pp.115-121
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• 1989

A survey on the prevalence and distribution of antibodies to BLV was performed by the agar-gel immunodiffusion test over a period from 1983 to 1985. More than 2,407 serum samples were collected from Holstein cattle raised in the eastern part of Saitama prefecture where suburban dairy farm is operated. The average positive rate of this period was 4.9%. The rates of reactive samples varied from 2.6 to 9.8% among the age groups of cattle from younger than one year to 14 years of age. The positive rate increased gradually with age. The positive rates also varied widely from 0 to 21% among areas surveyed. Furthermore, there were large differences in this rate among farms even in the same area. The results were interpreted and discussed in connection with the enzootic feature of BLV infection.

• The Korean Journal of Zoology
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• v.39 no.3
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• pp.307-316
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• 1996

A persistent major haemolyruph protein (MHP) was confirmed, and its ontogeny and physicochemical charadedstics were investigated in Helicoverpa assulta. The MHP existed continually during larval-pupal-adult development, and its ontogeny was similar to that of total haemolymph protein concentration during development. Its content increased with larval growth, and kept to high level during pupal-adult development except for temporary decrease at the early pupal and adult stages. The MHP was purified by ammonium sulfate precipitation, gel filtration and ion exchange chromatography. The purified MHP was determined to be hexamer glycolipoprotein (pI 5.9, M.W. 414kDa) consisted of single type subunit (69kDa). Amino acid analysis suggested that the MHP contained a relatively high content of aromatic amino acids (18.27 mole % of tryptophan, 7.47 mole % of tyrosine and 6.51 mole % of phenylalanine) compared to storage proteins from other insects. Immunodiffusion test and electrophoretic analysis of the organ proteins (gut, fat body, and Malphigian tubule) suggested that the major haemolymph protein was present in the fat body.

• Korean Journal of Veterinary Service
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• v.17 no.2
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• pp.95-113
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• 1994

To establish an effective diagnostic measure for detection of the antibodies against Actinobacillus pleuropneumoniae, the methods for tube agglutination test (TAT), plate agglutination test (PAT), micro-agglutination test(MAT) and agar-gel immunodiffusion test(ID) were improved and standarized, and the comparative studies were carried out. The results obtained through the experiments were summarized as follows. 1. The rabbit hyperimmune sera to reference serotypes 1 to 6 were cross-tested with TAT, PAT, MAT and ID. In the homologous systems, the range of antibody titers in TAT was 80 to 640, showing the cross-reaction in serotypes 3, 4, 5 and 6. The range of antibody titers in PAT was 4 to 64, showing the cross-reaction in serotypes 3, 4, 5 and 6. In ID, the range of antigen titers was 8 to 32, and cross-reaction was observed in serotype 5. 2. The optimal concentration of antigen in PAT and MAT were 100mg /ml and 1.25mg /ml respectively. The most sensitive reaction in MAT was observed in 52$^{\circ}C$ for 18hrs. 3. In ID, the most promising antigen and the buffer for agar-gel were EDTA-treated antigen and 0.05M tris buffer (pH 7.2), respectively. 4. By the tests for 200 swine sera, it was found that the frequency of positive reaction were 203 in TAT, 240 in PAT and 163 in ID. 5. When compared the titers of TAT with those of MAT for 200 swine sera, MAT showed the higher titer than TAT being increased by relative correlation. Int was found that the titer for positive readings were 20 in TAT and 40 in MAT. 6. when compared the results of ID with those of TAT for 200 swine sera, all sera with TAT titer under 10 were negative in ID. Of the sera with TAT titer 20 and 40, 55.1% nd 91.8% were positive in ID, respectively. All sera with TAT titer above 80 were positive in ID. In comparison of ID and MAT, all sera with MAT titer under 20 were negative in ID. Of the sera with MAT titer 40 and 80, 24.7% and 93.9% were positive in ID, respectively. All sera with MAT titer over 160 showed positive in ID. 7. In conclusion, the established MAT showed high sensitivity but low specificity, wherease ID revealed low sensitivity but high specificity.

• Korean Journal of Veterinary Service
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• v.19 no.3
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• pp.221-226
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• 1996

IBD's antibody level and morphological change of immune organ was examined in chicken. The results were as follows ; 1. Seventy percent at 42 day and 40% at 45 day of age chickens were reacted positively by the agar gel immunodiffusion test and 42 day and 45 day of age chickens indicated 1859, 1425 by the ELISA test, respectively. 2. In 2 and 5 day young broiler chickens, the level of maternal antibody was not proper. B/B ratio showed low level, but S/B ratio and BS were normal. 3. In layer, 100% of 8 and 86 day of age chicken and 70% at 40 day of age chicken had antibody aganist IBDV. The level of antibody was high as 2293 and 3336 in 8 and 86 day of age chicken, while was low as 1186 in 40 day of age chicken. B/B ratio showed low level and S/B ratio high level, but BS was normal.