효소면역법을 이용한 Brucella abortus 항체 검출에 관한 연구

심항섭 ( Hang Sub Shim ),국정희 ( Jung Hee Kook ),정봉수 ( Bong Su Chung ),고태오 ( Tae Oh Ko ),조중현 ( Jung Hyun Cho ),박유순 ( Yu Soon Park ) 한국동물위생학회 1998 한국동물위생학회지 (KOJVS) Vol.21 No.2

In order to establish a rapid, sensitive and specific diagnostic method for detection of antibody to Brucella abortus, a enzyme-linked immunosorbent assay(ELISA) was adapted. The diagnostic efficacy of the established ELISA was compared with that of the standard tube agglutination test for B abortus. 1. It was found that the optimal concentration of antigen for this ELISA was 51ug/m1, the optimal dilution of conjugate was 1:2000, and the optimal dilution of serum was 1:200, respectively. 2. Cut off value in this ELISA was 1,102 that was determined by mean absorbance(at 492nm) of tube agglutination test negative serum added with the triple value of the standared devation. 3. The relationship between the tube agglutination test and ELISA was showen high corresponding rate with sensitivity(96.3%) and specificity(98.1%). 4. The efficacy of the ELISA for detection of B abortus antibody was compared with tube agglutination test in brucellosis outbreak farm. The sensivity of ELSIA was higher than tube agglutination test.

Short communication : Tube agglutination test is superior than other serological tests for diagnosis of brucellosis in small ruminants

( Md. Siddiqur Rahman ),( Nusrat Jahan ),( Mohammad Arif Hossain ),( M. J. Uddin ),( Niraj Kanti Shil ),( KBM Saiful Islam ),( Md. Shamim Ahasan ),( A. K. M. Anisur Rahman ),( Hee Jong Song ) 한국동물위생학회 2008 한국동물위생학회지 (KOJVS) Vol.31 No.4

Brucella spp. are small, non-motile Gram-negative coccobacilli known to cause disease in a number of vertebrate species including humans and brucellosis is one of the world`s major zoonoses, alongside bovine tuberculosis and rabies. There are about 33.55 million goats and 1.16 million sheep in Bangladesh. The sheep and goats can significantly play an important role in the economic well being of the resource-poor farmer in Bangladesh. Sexually matured 362 female small ruminants (300 goats and 62 sheep) were examined. Approximately 3-5ml of blood was collected from the jugular vein of each animal and sera samples were prepared. Samples were then tested for brucellosis by using Rose Bengal test(RBT), plate agglutination test(PAT) and tube agglutination test(TAT). Among 362 small ruminants, irrespective of species(sheep or goat), diagnosed highest in TAT, 2.21%(n=8) and lowest both by RBT & PAT, 1.93%(n=7) and it is concluded that TAT is superior than RBT and PAT.

Interpretation of tube agglutination test for bovine brucellosis with turbidimetric readings and international unit

( Dong Hee Cho ),( Hyang Mi Nam ),( Jong Wan Kim ),( Eun Jeong Heo ),( Yun Sang Cho ),( In Yeong Hwang ),( Jae Myung Kim ),( Jong Man Kim ),( Suk Chan Jung ) 한국동물위생학회 2007 한국동물위생학회지 (KOJVS) Vol.30 No.2

The tube agglutination test has been used for bovine brucellosis diagnosis in Korea since middle 1950s. The reported high specificity was its value in eradication program. However, the reading of reaction mostly depends on personal experience, thus here we report a way to improve accuracy and uniformity of reading. The tube agglutination was conducted according to the protocol provided by Korean Ministry of Agriculture and Forestry. The intensity of reaction was measured by spectrophotometer. The relationship between turbidity and percentage clearing was generally in direct proportion and linear. The correspondent percent transmittance at 75, 50, and 25% clearing were 91, 82, and 73, respectively. Then, the degree of percentage of clearing at given international unit was measured. With about 1.5 unit of serum, the maximum percentage clearing was observed. The international unit showing 25, 50, and 75 percentage clearing were 0.61, 0.83 and 1.35, respectively. Based on the information obtained using international standard serum, the calculation of international unit of test serum was available. According to the protocol for bovine brucellosis diagnosis which provided by Korean Ministry of Agriculture and Forestry, the available range of detectable international unit was between 15 and 538. And the corresponding international units for suspicious case ranged between 42 and 127. Of the 35 sera from B abortus infected cattle, about half of them had more than 538 international units. Collectively, the reading of turbidity using spectrophotometer and application to international unit improved accuracy and uniformity of reading.

Interpretation of tube agglutination test for bovine brucellosis with turbidimetric readings and international unit

Cho, Dong-Hee,Nam, Hyang-Mi,Kim, Jong-Wan,Heo, Eun-Jeong,Cho, Yun-Sang,Hwang, In-Yeong,Kim, Jae-Myung,Kim, Jong-Man,Jung, Suk-Chan The Korean Society of Veterinary Service 2007 韓國家畜衛生學會誌 Vol.30 No.2

The tube agglutination test has been used for bovine brucellosis diagnosis in Korea since middle 1950s. The reported high specificity was its value in eradication program. However, the reading of reaction mostly depends on personal experience, thus here we report a way to improve accuracy and uniformity of reading. The tube agglutination was conducted according to the protocol provided by Korean Ministry of Agriculture and Forestry. The intensity of reaction was measured by spectrophotometer. The relationship between turbidity and percentage clearing was generally in direct proportion and linear. The correspondent percent transmittance at 75, 50, and 25% clearing were 91, 82, and 73, respectively. Then, the degree of percentage of clearing at given international unit was measured. With about 1.5 unit of serum, the maximum percentage clearing was observed. The international unit showing 25, 50, and 75 percentage clearing were 0.61, 0.83 and 1.35, respectively. Based on the information obtained using international standard serum, the calculation of international unit of test serum was available. According to the protocol for bovine brucellosis diagnosis which provided by Korean Ministry of Agriculture and Forestry, the available range of detectable international unit was between 15 and 538. And the corresponding international units for suspicious case ranged between 42 and 127. Of the 35 sera from B abortus infected cattle, about half of them had more than 538 international units. Collectively, the reading of turbidity using spectrophotometer and application to international unit improved accuracy and uniformity of reading.

Interpretation of tube agglutination test for bovine brucellosis with turbidimetric readings and international unit

정석찬,조동희,남향미,김종완,허은정,조윤상,황인영,김재명,김종만 한국동물위생학회 2007 韓國家畜衛生學會誌 Vol.30 No.2

The tube agglutination test has been used for bovine brucellosis diagnosis in Korea since middle 1950s. The reported high specificity was its value in eradication program. However, the reading of reaction mostly depends on personal experience, thus here we report a way to improve accuracy and uniformity of reading. The tube agglutination was conducted according to the protocol provided by Korean Ministry of Agriculture and Forestry. The intensity of reaction was measured by spectrophotometer. The relationship between turbidity and percentage clearing was generally in direct proportion and linear. The correspondent percent transmittance at 75, 50, and 25% clearing were 91, 82, and 73, respectively. Then, the degree of percentage of clearing at given international unit was measured. With about 1.5 unit of serum, the maximum percentage clearing was observed. The international unit showing 25, 50, and 75 percentage clearing were 0.61, 0.83 and 1.35, respectively. Based on the information obtained using international standard serum, the calculation of international unit of test serum was available. According to the protocol for bovine brucellosis diagnosis which provided by Korean Ministry of Agriculture and Forestry, the available range of detectable international unit was between 15 and 538. And the corresponding international units for suspicious case ranged between 42 and 127. Of the 35 sera from B abortus infected cattle, about half of them had more than 538 international units. Collectively, the reading of turbidity using spectrophotometer and application to international unit improved accuracy and uniformity of reading.

Tube agglutination test is superior than other serological tests for diagnosis of brucellosis in small ruminants

Rahman, Md. Siddiqur,Jahan, Nusrat,Hossain, Mohammad Arif,Uddin, M.J.,Shil, Niraj Kanti,Islam, KBM Saiful,Ahasan, Md. Shamim,Rahman, A.K.M. Anisur,Song, Hee-Jong The Korean Society of Veterinary Service 2008 韓國家畜衛生學會誌 Vol.31 No.4

Brucella spp. are small, non-motile Gram-negative coccobacilli known to cause disease in a number of vertebrate species including humans and brucellosis is one of the world's major zoonoses, alongside bovine tuberculosis and rabies. There are about 33.55 million goats and 1.16 million sheep in Bangladesh. The sheep and goats can significantly play an important role in the economic well being of the resource-poor farmer in Bangladesh. Sexually matured 362 female small ruminants(300 goats and 62 sheep) were examined. Approximately 3-5 ml of blood was collected from the jugular vein of each animal and sera samples were prepared. Samples were then tested for brucellosis by using Rose Bengal test(RBT), plate agglutination test(PAT) and tube agglutination test(TAT). Among 362 small ruminants, irrespective of species(sheep or goat), diagnosed highest in TAT, 2.21%(n=8) and lowest both by RBT & PAT, 1.93%(n=7) and it is concluded that TAT is superior than RBT and PAT.

돼지에서 Actinobacillus pleuropneumoniae의 혈청학적 진단법에 대한 비교연구

심항섭 ( Hang Sub Shim ),우종태 ( Jong Tae Woo ),조중현 ( Jung Huin Cho ),전무형 ( Moo Hyung Jun ) 한국동물위생학회 1994 한국동물위생학회지 (KOJVS) Vol.17 No.2

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 l.25mg /ml respectively. The most sensitive reaction in MAT was observed in 52°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.

ABO 항체 역가 측정과 교차시험의 Column Agglutination Technique과 Tube Test 비교

임영애 대한수혈학회 2023 大韓輸血學會誌 Vol.34 No.3

Background: ABO Ab titration using Erytra Eflexis (Diagnostic Grifols, Barcelona, Spain, abbreviated as Eflexis) and a pretransfusion crossmatching using DG Gel Coombs Card (Diagnostic Grifols, abbreviated as Card) were compared with the tube test (TT) to help laboratories. Methods: Samples subjected to TT were titrated for ABO antibody using the Eflexis. The titers obtained were thereafter converted into log steps of 2 (n=20 for each ABO group). The reference range (RR) of the ABO Ab titer using Eflexis was determined using samples from health checkup patients (n=30 for each ABO group). The Card and LISS-antihuman globulin (AHG) test of TT were compared for 40 cases with identified antibodies. Results: The average titers of IgM anti-B of blood group A by TT (3.2 vs 2.4, P<0.001), and anti-A of type O by Eflexis (4.5. vs 5.5, P=0.001 for IgM, 6.3. vs 7.3, P<0.001 for total antibody) were higher compared to the alternative method. The RRs of IgM (total antibody) of anti-B and anti-A were determined to be 1:2≤ (1:2≤) for anti-B in group A, 1:8≤ (1:8≤) for anti-A in group B, and 1:8≤ (1:32≤) and 1:16≤ (1:32≤) for anti-B and anti-A in group O, respectively. A comparison of crossmatching revealed that the average hemagglutination strength of the Card was 1.65, which was stronger than the LISS-AHG of 0.50 (P<0.001). Conclusion: The results of this study are expected to provide valuable insights for the establishment and guidance of laboratory tests utilizing Eflexis and Card. (Korean J Blood Transfus 2023;34:189-197)

소 브루셀라병의 혈청학적 진단법 비교실험

허진 ( Jin Hur ),( Ibulaimu Kakoma ),정재명 ( Jae Myong Jeong ),이현진 ( Hyeon Jin Lee ),백병걸 ( Byeong Kirl Baek ) 한국동물위생학회 2007 한국동물위생학회지 (KOJVS) Vol.30 No.3

A novel enzyme linked immunosorbent assay(ELISA) is described and compared with other established serologic tests for bovine brucellosis, namely the rose bengal test(RBT), the complement fixation test(CFT), and the tube agglutination test(TAT) approved and used in Korea. A total of 109 bovine serum samples were tested using all the 4 assays and analyzed as to specificity, sensitivity, reproducibility and predictive value. The ELISA showed 100% agreement with the CFT. The least agreement between ELISA was observed with the TAT. The agreement between the ELISA and the RBT was not significantly different from that observed between the CFT and the ELISA. It is concluded that the new assay would be a good candidate for routine serologic survey for brucellosis in Korea. A protocol combining the ELISA and the CFT would increase the power for detection of serologically positive individuals and herds.

소 브루셀라병 진단을 위한 라텍스 응집반응 시험법

백병걸,허진,진찬문 한국수의공중보건학회 1999 예방수의학회지 Vol.23 No.3