• Title/Summary/Keyword: Brucella abortus

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DNA fingerprinting of Brucella abortus isolated from bovine brucellosis outbreaks by repetitive element sequence (rep)-PCR

  • Suh, Dong Kyun
    • Korean Journal of Veterinary Research
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    • v.45 no.2
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    • pp.199-205
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    • 2005
  • DNA fingerprint patterns of 8 Brucella reference strains and 15 B. abortus field isolates were characterized by repetitive element sequence-based PCR (rep-PCR) using BOX- and ERIC-primers in this study. AMOS PCR differentiated all Brucella field isolates from B. abortus RB51, a vaccine strain by producing a B. abortus-specific 498 bp band. Rep-PCR using BOX-primer produced 13 to 18 bands with sizes of between 230 and 3,300 bp, and discriminated Brucella strains to the species level except B. canis and B. suis. PCR products amplified with ERIC primers were, however, not appropriate for differentiating the Brucella isolates. DNA fingerprint patterns for all B. abortus field isolates were identical among them and were put on one cluster with B. abortus biovar 1 reference strain in the dendrogram, indicating they were highly clonal. These results suggested that rep-PCR using BOX primer might to be a useful tool for calculating genetic relatedness among the Brucella species and for the study of brucellosis epidemiology.

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
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    • v.21 no.2
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    • pp.107-115
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    • 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.

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Biochemical characteristics of Brucella abortus isolated from cattle in Gyungbuk province (경북지방 소에서 분리한 Brucella abortus의 생화학적 특성)

  • Kim, Seong-Guk;Kim, Young-Hoan;Cho, Min-Hee;Lee, Young-Ju;Park, Cheong-Kyu
    • Korean Journal of Veterinary Service
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    • v.32 no.2
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    • pp.139-146
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    • 2009
  • Bovine brucellosis is a zoonosis, long incubation period and chronic infectious disease, usually caused by Brucella abortus. This study was carried out to investigate the biotyping and biochemical characterization of B. abortus isolated from 208 farm 871 korean cattle and holstein diagnosed brucellosis by serological positive in Gyeongbuk province during the period from 2002 to 2006. B. abortus was isolated from 124 (14.2%) of 871 cattle, and isolated 110 (13.4%) of 820 Korean cattle and 14 (27.5%) of 51 holstein in breed. The uterus of korean cattle was isolated in 8 (17.8%) of 45 cattle and supramammary lymph none of holstein was isolated 11 (68.8%) of 16 cattle. 101 (12.5%) of 810 serological positive blood samples were isolated B. abortus. The isolation rate of B. abortus was correlated with antibody titers. The biochemical characterization of isolates was non-hemolytic, production of H$_2$S, oxidase-positive, catalase-positive, hydrolyzation of urea and growth of basic fuchsin dye medium. As a result, all of isolates was identified B. abortus bv 1. 124 isolates were susceptible to ampicillin, lincospectin, amikacin, gentamicin, kanamycin, neomycin, streptomycin, tetracycline, ciprofloxacin, norfloxacin and enrofloxacin.

Development of ELISA for Brucella abortus RB51 II. Purification of 8kDa antigen and development of ELISA using its antigen of Brucella abortus RB51 (부루세라 RB51의 ELISA 진단법 개발 II. Brucella abortus RB51균의 8kDa 항원 정제 및 ELISA 진단법 개발)

  • Her, Moon;Cho, Dong-hee;Jung, Byeong-yeal;Cho, Seong-kun;Jung, Suk-chan;Kim, Ok-kyung
    • Korean Journal of Veterinary Research
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    • v.41 no.1
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    • pp.51-57
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    • 2001
  • A procedure for extraction and purification of 8 kDa antigen of Brucella abortus RB51 was developed. Bacteria heat inactivated at $60^{\circ}C$, 30 min was extracted by 1% sarcosine and followed by fluid pressure liquid gel filtration chromatography of 2 series, Superose 12 HR 10/30 and Sephacryl S-100. There was produced $71.46{\mu}g/g$(wet) of 8 kDa antigen, and it resisted 1% trypsin, solved 1% triton X-100 higher than distilled water and inactivated 0.1% proteinase K. These results show that 8 kDa antigen may be a lipoprotein existed cell surface of B. abortus RB51. Also, we developed ELISA using purified 8 kDa surface antigen of Brucella abortus RB51 strain, its specificity and sensitivity was 95.0%, 98.6%, respectively. As compared with dot-blot assay using whole cell and ELISA using 8 kDa antigen, its correlation was 93.5%.

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Serological cross-reaction with Brucella abortus antigen extracted by sodium dodecyl sulfate and Yersinia enterocolitica 0:9 (SDS 처리한 브루셀라 항원과 Yersinia enterocolitica 0:9주의 혈청학적 교차반응 연구)

  • Lim, Yoon-kyu;Yang, Ki-chun;Lee, Kyung-kap;Park, Jun-hong;Lee, Du-sik;Park, Yong-ho;Kang, Seung-won;Mok, Ji-won;Lee, Yong-soon
    • Korean Journal of Veterinary Research
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    • v.35 no.1
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    • pp.143-148
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    • 1995
  • Brucella abortus cell wall antigen was extracted from Brucella abortus 1119-3 by ultrasonication and followed by sodium dodecyl sulfate(SDS) treatment. In order to confirm whether this preparation is serologically cross reactive with Yersinia enterocolitica 0:9, Western blot analysis with mouse anit-Brucella abortus1119-3 and with mouse anti-Yersinia enterocolitica 0:9 was performed. ELISA results from using those Brucella antigen and Yersinia antigen were assessed whether they had correlation. According to the results of western blot analysis and ELISA, there was no evidence of cross reactivity between the Brucella abortus 1119-3 antigen preparation and Yersinia enterocolitica 0:9. Therefore the SDS treated antigen prepared in this study could be suitably used as specific ELISA antigen without confusion in the interpretation of serological tests for brucellosis in cattle.

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Genotyping of Brucella abortus isolated from cattle in Gyeongbuk province by MLVA (MLVA를 이용한 경북지역 소에서 분리된 Brucella abortus의 유전형별)

  • Kim, Seong-Guk;Kim, Young-Hoan;Lee, Hong-Young;Choi, Jeong-Hye;Choi, Seong-Kyoon
    • Korean Journal of Veterinary Service
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    • v.34 no.3
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    • pp.227-234
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    • 2011
  • Brucella (B.) abortus is a facultative intracellular pathogen that infects a wide variety of animal species and human. Brucellosis is the zoonosis and an extremely important disease around the world. Although the eight species can be differentiated by conventional phenotypic tests, these species display a high degree of DNA homology in DNA-DNA hybridization assay (>90%). Various methods have been established for genotyping of Brucella species, but most of analytical methods are lack reproducibility and limited capability to differentiate them. The attempt of this study was to evaluate multiple-locus VNTR analysis (MLVA) for use of epidemiological trace-back analysis in bovine brucellosis. Ninety-four B. abortus isolates from Gyeongbuk province during 2006~2010 were analyzed using 16 VNTR locus. High resolution automatic capillary electrophoresis system was used for more throughput, simpleer, faster, and better discriminable than conventional gel electrophoresis. As a result, 13 different genotypes were identified from 94 B. abortus isolates. MLVA could contribute to epidemiological trace-back analysis of bovine brucellosis.

Different invasion efficiencies of Brucella abortus wild-type and mutants in RAW 264.7 and THP-1 phagocytic cells and HeLa non-phagocytic cells

  • Shim, Soojin;Im, Young Bin;Jung, Myunghwan;Park, Woo Bin;Yoo, Han Sang
    • Korean Journal of Veterinary Research
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    • v.58 no.2
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    • pp.95-98
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    • 2018
  • Brucellosis is one of the common zoonoses caused by Brucella abortus (B. abortus). However, little has been reported on factors affecting invasion of B. abortus into host cells. To investigate cell-type dependent invasion of B. abortus, phagocytic RAW 264.7 and THP-1 cells and non-phagocytic HeLa cells were infected with wild-type and mutant B. abortus, and their invasion efficiencies were compared. The invasion efficiencies of the strains were cell-type dependent. Wild-type B. abortus invasion efficiency was greater in phagocytic cells than in epithelial cells. The results also indicated that there are different factors involved in the invasion of B. abortus into phagocytic cells.

Isolation and biotyping of Brucella abortus from dairy cattle in Kyungpook area, Korea (경북지방(慶北地方) 젖소로 부터 Brucella abortus의 분리(分離) 및 균형별(菌型別))

  • Chung, Jong-sik;Cho, Yoong-jun;Park, Cheong-kyu
    • Korean Journal of Veterinary Research
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    • v.28 no.2
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    • pp.339-343
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    • 1988
  • Serological study and isolation of Brucellae from dairy cattle were carried out in Kyungpook area of Korea during the period from 1984 to 1985. Biotyping of the isolates was done. Of the 11168 cattle in 2479 herds examined, 27 cattle (0.24%) in the 17 herds (0.69%) were serologically positive for brucellosis. Brucella abortus was recovered from 20 (74.1%) of the 27 infected cattle, and all the strains of the isolates belonged to biotype 1. Of the tissues cultured for brucellae, the supramammary lymph nodes had the highest rate (55.6%) of recovery of brucella organisms.

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The virulence of Brucella abortus isolated from cattle in Korea (국내 소에서 분리한 Brucella abortus의 병원성 분석)

  • Lim, Jeong Ju;Kim, Jeong-Hwa;Kim, Dong Hyeok;Lee, Jin Ju;Kim, Dae Geun;Jun, Moo-Hyung;Kim, Sang Hun;Chang, Hong Hee;Lee, Hu Jang;Min, Won-Gi;Kim, Suk
    • Korean Journal of Veterinary Research
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    • v.51 no.1
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    • pp.15-20
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    • 2011
  • In this study, we isolated 12 of Brucella (B.) spp. from cattle, which have been positive in Rose Bangal test and tube agglutination test in Gyeongbuk province in 2009. According to AMOS PCR analysis, isolated 12 strains were identified as B. abortus. Murine derived macrophage, RAW 264.7 cells, were infected with isolated 12 strains or reference strain (B. abortus 544), and bacterial internalization were characterized. According to these results, we divided the isolated strains into the following three groups: class I, lower internalization than that of B. abortus 544; class II, similar internalization to that of that of B. abortus 544; class III, higher internalization than that of B. abortus 544 within RAW 264.7 cells. Furthermore, intracellular growth, bacterial adherent assay, LAMP-1 colocalization, virulence in mice and surface protein pattern were characterized. From these results, representative strains of class III showed lower LAMP-1 colocalization, higher adherent efficiency, higher virulence in mice than those of B. abortus 544, and showed different pattern of surface proteins. These results suggest that B. abortus field strains, isolated from cattle in Korea, possess various virulence properties and higher internalization ability of field strain may have an important role for its virulence expression.