• Title/Summary/Keyword: Tube agglutination test

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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
    • Korean Journal of Veterinary Service
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    • v.30 no.2
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    • pp.243-249
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    • 2007
  • 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.

Comparative Studies on Serological Tests for Actinobacillus pleuropneumoniae Infection in Swine (돼지에서 Actinobacillus pleuropneumoniae의 혈청학적 진단법에 대한 비교연구)

  • 심항섭;우종태;조중현;전무형
    • 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.

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Evaluation of ABO Antibody Titration Using Tube and Column Agglutination Techniques (ABO 동종 응집소 역가 측정을 위한 시험관 및 미세원주응집법의 평가)

  • Cho, Chi-Hyun;Kim, Ha-Nui;Yun, Seung-Gyu;Choi, Gye-Ryung;Choi, Jae-Yeoul;Kim, Jang-Su;Lim, Chae-Seung;Kim, Young-Kee;Lee, Kap-No
    • Laboratory Medicine Online
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    • v.1 no.1
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    • pp.57-63
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    • 2011
  • Background: ABO antibody titration is useful for the evaluation of ABO-incompatible bone marrow or solid organ transplantations, yet the results quite vary between different test methods used. We compared the results of microcolumn agglutination and tube methods. Methods: Anti-A and anti-B isoagglutionin titers were determined in 63 healthy individuals (23 O, 20 A, and 20 B blood groups) using 4 different methods: immediate spin tube (tube), microcolumn agglutination without anti-human globulin (AHG) (CAT), tube with AHG (tube-AHG) and microcolumn agglutination with AHG (CAT-AHG). Results: The median (range) titers of anti-A and anti-B in group O individuals by tube, CAT, tube-AHG, and CAT-AHG methods were 64 (8-512), 64 (8-512), 128 (8-2,048), and 128 (16-2,048); 64 (16-128), 128 (16-256), 128 (16-512), and 256 (16-512), respectively. The median (range) titers of anti-A in group B and anti-B in group A individuals by the four methods were 64 (16-128), 128 (8-128), 128 (8-256), and 256 (8-256); 64 (8-128), 64 (8-128), 32 (8-128), and 64 (8-256), respectively. The isoagglutinin titer measured by CAT-AHGmethod was the highest. The titers measured by CAT and CAT-AHG methods were 0-1 titer higher than those by tube and tube-AHG methods, respectively. Whatever method was used, the isoagglutinin titers were higher in women than in men. Conclusions: CAT-AHG was the most sensitive method among the four methods tested. Since AHG titer values are critical for the clinical management and CAT has less manual procedures than tube method, CAT-AHG method could be used for the standardization of ABO antibody titration in different institutions.

Comaprison of Six Serological Methods for the Diagnesis of Bovine Brucellosis (브루셀라병(病) 검색(檢索)에 사용(使用)되는 여러가지 혈청진단법(血淸診斷法)의 비교연구(比較硏究))

  • Kim, Keum-Hwa;An, Soo-Hwan;Park, Yong-Ho;Kim, Dong-Sung
    • Korean Journal of Veterinary Research
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    • v.22 no.2
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    • pp.149-153
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    • 1982
  • Results obtained from six secological tests for diagnosing bovine brucellosis-standard plate agglutination test (SPT), standard tube agglutination test(STT), complement fixation test(CFT), Rivanol test (RT), agar gel precipitation test (AGP) and counterimmunoelectrophoresis(CIEP) were compared using 38 sera from brucella reactors and 222 sera from dairy and beef cattle in field. The SPT gave 1.6% apparent false negative reactions and 15.4% apparent false positive reactions when compared with STT which is an official test for bovine brucellosis in this country. The distribution of antibody titers determined by STT showed that 37.5% of 38 reactors had antibody titers ranging from 100 to 200, and the remaining 62.5% had antibody titers of 400 or higher. when 38 reactor judged by STT were tested by CFT and RT, 32 cattle(82.4%) were positive by CFT and 33 cattle (86.8%) were positive by RT, respectively. This results suggest that RT is comparable to CFT in the diagnosis of bovine brucellosis. The results also indicated that both AGP and CIEP were insensitive to detect brucella infection in cattle.

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Isolation of Anaplasma centrale from Korean Cattle (아나플라즈마병(病)에 관(關)한 연구(硏究) 제(第) II 보(報) 한우(韓牛)에서 Anaplasma centrale 분리(分離))

  • Jeon, Yeong
    • Korean Journal of Veterinary Research
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    • v.18 no.1
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    • pp.19-22
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    • 1978
  • In order to isolate and identify Anaplasma bodies, two dairy cattle were inoculated with the whole blood of Korean cattle reacted to capillary tube agglutination test (Ana-test). The results obtained are as follows: 1. In the infected dairy cattle, Anaplasma bodies were first detected between 30 to 40 days post infection. 2. The isolates were identified as Anaplasma centrale on the basis of morphology of infected anaplasma bodies in erythocytes.

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Serological monitoring on brucellosis in livestock of Korea (국내 가축에서 브루셀라병에 대한 혈청학적 모니터링)

  • Sung, So-Ra;Kim, Ji-Yeon;Her, Moon;Lee, Kichan;Kang, Sung-Il;Lee, Hyang-Keun;Cho, Hyo Rim;Lee, Jin Ju;Jung, Suk Chan
    • Korean Journal of Veterinary Research
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    • v.54 no.4
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    • pp.197-201
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    • 2014
  • In Korea, brucellosis has been reported periodically in cattle and rarely in dogs; however, it has not previously been screened in domestic animals such as elk, pigs and goats. To investigate the serological prevalence, serum samples were taken from the aforementioned animals annually during 2007-2013 and screened by the rose-bengal test (RBT) or modified RBT, after which positive sera were evaluated by the standard tube agglutination test (STAT). Finally, RBT and STAT-positive sera were confirmed by competitive-ELISA. Brucella abortus biovar 1 was isolated from three elk that were shown to be positive serologically in 2008. There was no evidence of brucellosis in pigs. Based on serological monitoring and investigation of etiological agents, there is no evidence of outbreak of brucellosis in elk, pigs or goats of Korea since 2008. However, the possibility for brucellosis from cattle to affect these other livestock exists; therefore, extensive and continuous serological monitoring is required to maintain their brucellosis-free status.

Serological survey of diseases to poultry and swine in slaughtered ostriches (도축 타조에서 닭 및 돼지 질병에 대한 혈청학적 조사)

  • Kim Soon-Tae;Park In-Hwa;Kim Young-Hoan;Cho Kwang-Hyun;Oh Kyu-Shil;Son Jae-Kweon;Jyeong Jong-Sik
    • Korean Journal of Veterinary Service
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    • v.27 no.3
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    • pp.281-288
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    • 2004
  • As all other intensively farmed domestic species, most mortality in ostriches is closely to rearing conditions. While ostriches is also highly sensitive to stress, species-specific infectious disease play only a minor role. But investigation of ostrich's disease is not peformed almost in Korea. The study was performed to investigate the titers of antibody for Newcastle disease(ND), Infectious bronchitis(IB), Egg drop syndrome '76(EDS), Avian influenza(AI), salmonellosis, Mycoplasma gallisepticum infection(MG), Mycoplasma synoviae infection(MS), Infectious bursal disease(IBD), Brucellosis, Toxoplasmosis, Japanese encephalitis(JE), Porcine parvovirus infection, Encephalomyocarditis and Porcine reproductive respiratory syndrome (PRRS). The results obtained in the 62 ostrich sera slaughtered in Gyeongbuk province were summarized as follows: The average of antibody positive rates to ND, IB, EDS, AI(H9Nl), JE, Porcine parvovirus infection and Encephalomyocarditis by HI test were $75.8\%,\;100\%,\;0\%,\;0\%,\;51.6\%,\;50\%\;and\;56.5\%$ respectively. The antibody positive rates to salmonellosis, MG, MS by plate agglutination test were $12.9\%,\;25.8\%,\;and\;0\%$ respectively. Antibodies to disease agent such as IBD and AI by agar gel precipitation(AGP) test, Brucellosis by tube agglutination, toxoplasmosis by latex agglutination test and PRRS by IFA were all negative.

Development of ELISA for detection of canine brucellosis (Canine brucellosis 검출을 위한 ELISA 진단법 확립)

  • Hur, Jin;Baek, Byeong-Kirl
    • Korean Journal of Veterinary Service
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    • v.34 no.2
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    • pp.159-166
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    • 2011
  • This study was carried out to develope enzyme-linked immunosorbent assay (ELISA) for detection of canine brucellosis in dogs experimentally inoculated with Brucella abortus 1119-3 and B. canis RM666. Groups A, B and C of dogs (each group consisting of three dogs) were orally inoculated with approximately $5{\times}10^9$ colony-forming units of B. abortus and B. canis, and with sterile pyrogen-free PBS, respectively. The animals were monitored at regular intervals upto the 12th week post inoculation (PI) by standard tube agglutination test (STAT), plate agglutination test (PAT), Rose Bengal test (RBT), 2-mercaptoethanol rapid slide agglutination test (2ME-RSAT) and ELISA. The induced antibody titers in group A dogs were detected from the first week PI to the eighth week PI in STAT, PAT and RBT using the inactivated whole cells of B. abortus 1119-3 as antigens, while no sera in groups B and C dogs reacted with the antigens. In 2ME-RSAT using whole cells of B. canis M-strain as antigens, the induced antibody titers in group B dogs were observed at the second week PI and persisted for the 12th week PI, while sera of groups A and C dogs did not react with the whole cells. In ELISA using cytoplasmic fractions antigen of B. abortus 1119-3, the mean optical density of antibodies in groups A and B was detected from the first and second weeks PI, respectively, and persisted for 12th week PI, while sera of group C did not cross-react with the fractions antigen. However, in ELISA using the hot saline extracts of B. canis M- as an antigen, the induced antibody titers in only group B dogs were detected from second week PI and persisted for until the end of this study. These results indicate that the ELISA using B. abortus 1119-3 cytoplasmic fractions as antigens can be a good candidate for detection of brucellosis by B. abortus as well as B. canis in dogs.

Studies on incidence of Bovine Brucellosis in Cheju-Do (제주도내 축우 부루셀라병 발생상황 조사)

  • 김우택;이완수;김공식
    • Korean Journal of Veterinary Service
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    • v.14 no.2
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    • pp.104-109
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    • 1991
  • Tile present study was carried out to investigate the incidence of bovine brucellsis in Cheju-do during the period from 1985 to 1990. The results were summarized as follows. 1. In the total 239,238 cattles tested. 1180(0.49%) were positive by standard tube agglutination test during the period from 1985 to 1990. 2. The major causes of incidence on brucellosis was grazing with carriers and repeated incidence in a herd. 3. The 13 Brucella abortus biotype 1 isolated from 10(50%) of 20 cattles slaughtered on brucellosis in 1990.

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Serosurvey for antibodies against brucellosis in pigs (돼지 brucellosis에 대한 항체가 조사)

  • Hur, Jin;Baek, Byeong-Kirl
    • Korean Journal of Veterinary Service
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    • v.34 no.2
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    • pp.153-157
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    • 2011
  • In order to investigate serum antibodies for detection of brucellosis in pigs, a total of 1208 sera were tested by Rose Bengal test (RBT), the standard tube agglutination test (STAT) and competitive ELISA (cELISA). The sera were collected from pigs of Gyeonggi, Chungnam, Chungbuk, Jeonnam and Jeonbuk, provinces during the period 2002 to 2004. All the sera were screened by RBT, and were confirmed by STAT and cELISA. Among 1208 sera, 26 sera (2.2%) were positive in screening test. All the 26 positive sera were positive by STAT, while all the sera were negative by cELISA. On the basis of this study, farmed pigs may be exposed to Brucella species. Furthermore, these results suggest that establishment of diagnoses for detection of porcine brucellosis is necessary.