• Title/Summary/Keyword: standard tube agglutination test

Search Result 17, Processing Time 0.027 seconds

Studies on enzyme-linked immunosorbent assay(ELISA) for detection of antibody to Brucella abortus (효소면역법을 이용한 Brucella abortus 항체 검출에 관한 연구)

  • 심항섭;국정희;정봉수;고태오;조중현;박유순
    • Korean Journal of Veterinary Service
    • /
    • v.21 no.2
    • /
    • pp.107-115
    • /
    • 1998
  • 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 5$\mu\textrm{g}$/ml, 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.

  • PDF

Efficacy of Brucella abortus strain RB51 vaccine in Korean mongrel dogs against virulent strains of B. abortus biotype 1 and B. canis

  • Hur, Jin;Baek, Byeong-Kirl
    • Korean Journal of Veterinary Service
    • /
    • v.33 no.1
    • /
    • pp.29-35
    • /
    • 2010
  • This study was performed to test the hypothesis that Brucella abortus strain RB51 (SRB51) might protect Korean indigenous mongrel dog against challenge with either virulent B. abortus biotype 1 or B. canis. A total of 12 Korean mongrel dogs were divided into four groups (Group A, B, C and D). Dogs belonging to Group A and C were inoculated subcutaneously with $1{\times}10^9$ CFU of SRB51 in 1ml of sterile phosphate buffered saline (PBS). Dogs of Group B and D were inoculated subcutaneously with 1ml of sterile PBS as control. At 12 weeks post vaccination, dogs of Group A and B were challenged by oral inoculation of virulent strain of B. canis ($5.0{\times}10^9$ CFU) and dogs of Group C and D were challenged by oral inoculation of virulent strain of B. abortus biotype 1 ($4.4{\times}10^{10}$ CFU). The serum antibodies titers in all dogs were monitored at regular interval for eight weeks after challenge (AC) by standard tube agglutination test, plate agglutination test, rose bengal test, 2-mercaptoethanol rapid slide agglutination test and 2-mercaptoethanol tube agglutination test. No antibody titers in Group A and C was detected. Also, the challenge strains were not found from blood of all dogs of Group A and C from 1 week AC till the end of the experiment by culture and modified AMOS-PCR, whereas B. canis and B. abortus challenge strains were detected from blood of Group B and D, respectively. In addition, neither of two challenge bacteria was recovered from liver, spleen, kidneys, lymph nodes and reproductive tracts of Group A and C dogs after postmortem. However, B. canis and B. abortus challenge strains were isolated from these tissues of Group B and D, respectively. These data suggest that SRB51 could be a promising vaccine candidate for immunizing dogs to control canine brucellosis caused by B. canis or B. abortus.

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
    • /
    • v.22 no.2
    • /
    • pp.149-153
    • /
    • 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.

  • PDF

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
    • /
    • v.30 no.2
    • /
    • pp.243-249
    • /
    • 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.

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
    • /
    • v.54 no.4
    • /
    • pp.197-201
    • /
    • 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.

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

  • 김우택;이완수;김공식
    • Korean Journal of Veterinary Service
    • /
    • v.14 no.2
    • /
    • pp.104-109
    • /
    • 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.

  • PDF

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
    • /
    • v.34 no.2
    • /
    • pp.159-166
    • /
    • 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.

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

  • Hur, Jin;Baek, Byeong-Kirl
    • Korean Journal of Veterinary Service
    • /
    • v.34 no.2
    • /
    • pp.153-157
    • /
    • 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.

Evaluation on diagnostic efficiency of the standard tube agglutination test for bovine brucellosis (소 브루셀라병 표준시험관응집반응법 진단효율 평가)

  • Sung, So-Ra;Kim, Ji-Yeon;Her, Moon;Lee, Kichan;Gu, Jeong-Hui;Kang, Sung-Il;Lee, Hyang-Keun;Kim, Suk-Mi;Jung, Suk-Chan
    • Korean Journal of Veterinary Service
    • /
    • v.35 no.4
    • /
    • pp.269-273
    • /
    • 2012
  • A confirmatory serological test, the standard tube agglutination test (STAT) is evaluated for the diagnostic efficiency in brucellosis Korea. A total of 345 bovine samples were collected from regional veterinary branch under national brucellosis monitoring program from January 2010 to June 2012 in Korea. These samples were diagnosed as suspected serum and brucellosis positive by the Rose Bengal test (RBT) and the STAT, respectively. The STAT was compared and evaluated with three serological test such as the indirect-enzyme linked immunosorbent assay (I-ELISA), competitive-enzyme linked immunosorbent assay (C-ELISA) and fluorescence polarisation assay (FPA) prescribed for international trade by OIE. Among the 345 bovine serum samples, 302 (87.5%) were diagnosed as positive in the STAT, while 215 (62.3%), 223 (64.6%) and 194 (56.2%) serum samples were diagnosed as positive for brucellosis in the I-ELISA, C-ELISA and FPA, respectively. The STAT showed quite high positive results as compared with three prescribed tests of OIE. FPA, I-ELISA and C-ELISA have shown 60.6%, 64.9% and 67.2% correlation, respectively as compared to the STAT. However correlations of three prescribed tests ranged high 84.1~97.7%. Especially, correlation between I-ELISA and C-ELISA is quite high, 97.7%. These results suggest that the STAT has shown many false-positive reactions. Therefore, additional serological test, such as ELISAs and FPA, would be necessary to adopt as a confirmatory test in the national surveillance program of bovine brucellosis in Korea.

Seroprevalence of abortion and stillbirth inducing disease in Hanwoo, in Jeonbuk eastern area (전북 동부지역 한우에서의 유사산 유발 질병 항체가 조사)

  • Song, Ji-Min;Shon, Ku-Rye;Koh, Won-Seok;Lee, Jeong-Won
    • Korean Journal of Veterinary Service
    • /
    • v.37 no.3
    • /
    • pp.179-183
    • /
    • 2014
  • Reproductive losses in a herd can be huge. Fetal reabsorption or undefined infertility often remain undetected. Routine herds monitoring for exposure, controlling the introduction of potential agent carriers, appropriate biosafety procedures, and vaccination where possible are together the best security against abortion and stillbirth inducing disease. For biosecurity of local farms, we performed antibody titers of abortion and stillbirth related diseases such as bovine viral diarrhea virus (BVDV), infectious bovine rhinotracheitis virus (IBRV), Neospora caninum, Toxoplasma gondii and Campylobacter fetus subsp venerealis. The blood samples were collected from 500 female Hanwoo over 1 year old of 100 farms in Jeonbuk eastern area. Champhylobater serological test was evaluated by the standard tube agglutination test (STAT) and other pathogen's antibodies were detected by indirect-enzyme linked immunoassay (I-ELISA). The seroprevalence of abortion and stillbirth inducing disease were BVDV 72.4%, IBRV 13.0%, N. caninum 1.2%, T. gondii 10.4% and C. venerealis 0.6%, irrespectively.