• 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.

• 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 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 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.

• 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.

• Asian-Australasian Journal of Animal Sciences
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• v.9 no.4
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• pp.465-469
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• 1996

Pathological and virological investigations were conducted on suspected outbreaks of infectious bursal disease (IBD) in a broiler farm and five pullet-raising poultry farms of Mymensingh and Tangail districts of Bangladesh. About 80 to 100 percent chicks were affected at the age of 26 to 45 days and mortality varied from 20 to 30 percent in broilers and 40 to 80 percent in layer chicks. Signs, symptoms, gross and microscopic lesions were typical of acute IBD. Several isolates of virus could be obtained by embryo inoculation and the virus was diagnosed as infectious bursal disease virus (IBDV) by agar gel immunodiffusion test (AGID). The virus isolate belonged to the very virulent pathotype of IBDV causing 100 percent mortality in three weeks old chicks on experimental infection.

• Korean Journal of Veterinary Research
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• v.34 no.1
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• pp.93-97
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• 1994

A immunological survey of paratuberculosis in dairy and Korean native cattles was conducted by enzyme linked immunosorbent assay(ELISA), complement fixation test(CFT), agar gel immunodiffusion test (AGID) and intradermal skin test(ID). Over all prevalence of pararuberculosis in cattles was 6.7%(109/1633) by ID, 7.5(205/2719) by AGID, 9.3% (245/2641) by CFT and 13.4%(363/2719) by ELISA. Prevalence in dairy cattle was higher than that of Korean native cattle. Of 70 ELISA-positive cattle, 23(28.6%) and 48(68.6%) cattles were classified as positive in the AGID and positive or suspect in CFT, respectively. Of 92 ELISA-suspect cattle, 32(34.9%) and 48(52.2%) cattles were classified as AGID-positive and CFT-positive or suspect, respectively. It was concluded that paratuberculosis is widespread in cattle of Korea.

• Korean Journal of Veterinary Service
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• v.29 no.2
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• pp.89-96
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• 2006

Bovine viral leukosis is a viral disease of cattle characterized by the development of tumors in the lymphatic tissue. A female Holstein, 3-year-old, was submitted for diagnosis at the Diagnostic laboratory, Chonbuk National University. Clinical sign of the affected animal showed emaciation, enlargement of superficial lymph node and mild diarrhea. Remarkable lesions were enlargement of many internal lymph nodes. Histopathology revealed excessive neoplastic lymphoid cells characteristic of BVL infection. Subsequently, serums from all cattle were collected and serological examination was done where a 85% seropositive rate was detected using ELISA test. ELISA method showed a comparatively 75% higher detection rate than the agar gel immunodiffusion (AGID) test (85% vs 40%). Serologically positive cattle were variably detected in all ages from under 1 year to over 6 year of age. Hematological examination consistently showed leukocytosis and a differential lymphocytosis of seropositive cattle. Detailed comparative pathological and serological data diagnosed the presence of bovine viral leukosis.

• Korean Journal of Veterinary Service
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• v.18 no.2
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• pp.133-151
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• 1995

During 2 years from Octorber 1992 to April 1994, prevalence of general respiratory diseases and atrophic rhinitis in the pig herds located in the Western Chungnam was investigated, and isolation of B. bronchiseptica was attempted for the pigs manifested with the clinical signs of atrophic rhinitis(AR). The isolates were characterized and identified in aspects of biochemical properties, antigenicity, drug sensitivity and pathogenicity. The results obtained through the experiments are summarized as follows; 1. During 2 years of investigation, the overall prevalence of the general respiratory diseases in the pi8 herds in Western Chungnam was 35.3%, consisting of 35.1% in the pig farms and 38.8% in a slaughter house. The prevalence by age groups accounts for 9.2% in adults, 44.7% in rearings and 25.3% in sucklings. By farm size, The highest prevalence of 56.5% was observed in the smallest farm with 1 to 200 heads. 2. The prevalence of clinical cases of artrophic rhinitis was recorded by 12.7% in the group that is the sows and piglets vaccinated, 28.9% in the group that is the sows only vaccinated and 39.8% in the group of the non-vaccinated groups. In the slaughter house, 53(24.8%) of 214 pigs examined exhibit the AR lesions. 3. A total of 189 strains of B. bronchiseptica were isolated from the pig herds. Isolation rates were 12.6% in the group that is the sows and piglets vaccinated, 34.1% in the group that is the sows only vaccinated and 45.7% in the group of the non-vaccinated groups. Isolation rate in the specimen from the slaughter house was 93( 43.5% ) of 214 pigs examined. Of the AR-non-vaccinated group, the piglets aged bet- ween 61 to 90 days revealed the highest isolation rate of 58.5%. 4. The titers of antibody against B. bronchiseptica were measured by tube agglutination test. The group that is the sow and piglet-vaccinated showed the highest titer of 640-2, 560 in sow and 640longrightarrow5, 120 in piglet. The group that is the sows only-vaccinated revealed 640-2, 560 in sows and 640-1, 280 in piglets. Both of the vaccinated groups showed 100% positive reaction. The group of the non-vaccinated sho-wed relatively lower titer of 0-1, 280 in both of sows and piglets. The positive rate of the sera obtained from the slaughter house was 53.3% with the antibody titer of 0-1, 280. 5. Biochemical and serological properities of 189 isolates were very similar to those of the reference B. bronchiseptical phase I type, indicating that most of isolates are B. bronchiseptica phase I type. 6. In antimicrobial drug susceptibility, 87.3% of 189 isolates was susceptible to chloramphenicol, 79.9%, to amikacin, 64.6%, to cephalothin and less than 35.4% to others. 7. In agar-gel immunodiffusion and SDS-PAGE analysis, the isolates presented the identical antigenicity and protein profiles to the reference standard strains. 8. The whole cells and bacterial filtrates of the isolates were inoculated to guinea pigs and mice. The isolates showed the hish pathogenicity and dermonecrotoxiciy.

• Current Research on Agriculture and Life Sciences
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• v.1
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• pp.195-199
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• 1983

A microplate enzyme-linked immunosorbent assay (ELISA) for detecting antibodies to bovine leukemia virus(BLV) is described and compared its sensitivity with that of the agar gel immunodiffusion test (ID) with BLV glycoprotein (gp) antigen using 263 sera collected in Korea and Japan. There was 98.5 per cent agreement between ELISA and ID when ELISA value, the value of tested serum(T) was devided with that of standard negative seurm(N) after the value of control was eliminated from T and N (T-C/N-C), of 1.5 or greater was considered positive. One hundred and forty four (99.6%) of 145 sera which were positive by ID were greater than 1.5 by ELISA, and 115 (97.5%) of 118 sera which were negative by ID were less than 1.5 by ELISA. As a result, it suggest that the ELISA test using BLV-gp antigen provides a useful serological tool for the diagnosis of BLV infection.