• IMMUNE NETWORK
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• 제2권4호
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• pp.195-201
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• 2002

Background: IgE is closely related to the development of allergies. However, the poor relationship between the specific IgE level and the severity of allergic diseases suggests the possibility of functionally different IgE isoforms. With this in mind, rat basophilic leukemia (RBL)-2H3 activation was analyzed with each type of rat IgE for two parameters, exocytosis and IL-4 mRNA production. RBL-2H3 has been well documented in the rat mucosal mast cell line. Methods: RBL-2H3 cells sensitized with each kind of rat IgE was activated by cross-linking FcRI with B5 (monoclonal anti-rat IgE mouse IgG antibodies). The RBL-2H3 exocytosis was measured by analyzing the ${\beta}$-hexosaminidase level, and the level of IL-4 mRNA synthesis was analyzed using semiquantitative RT-PCR. Rat IgE, which was produced by a parasite infection (REP), was prepared using either Paragonimus westermani metacercariae (REP-PW) or Anisakis simplex third stage larvae (REP-AS). A rat IgE prototype of IR162 was prepared by a peritoneal injection of immunocytoma. Results: The level of exocytosis showed a linear relationship with the rat IgE concentration when REP-PW or REP-AS was applied. However, it exhibited a biphasic response with IR162. In addition, the time course of heating at $56^{\circ}C$ illustrated the similarity between REP-PW and REP-AS, which differed from that of IR162. In contrast, the level of IL-4 mRNA synthesis in the RBL-2H3 cells with IR162 was comparable to that of either REP-PW or REP-AS. Conclusion: These results suggest that functionally different rat IgE isoforms exists in RBL-2H3 exocytosis.

• Journal of Microbiology and Biotechnology
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• 제21권11호
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• pp.1101-1108
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• 2011

The rise in global energy demand has prompted researches on developing strategies for transforming coal into a cleaner fuel. This requires isolation of microbes with the capability to degrade complex coal into simpler substrates to support methanogenesis in the coal beds. In this study, aerobic bacteria were isolated from an Indian coal bed that can solubilize and utilize coal as the sole source of carbon. The six bacterial isolates capable of growing on coal agar medium were identified on the basis of their 16S rRNA gene sequences, which clustered into two groups; Group I isolates belonged to the genus Rhizobium, whereas Group II isolates were identified as Chelatococcus species. Out of the 4 methods of whole genome fingerprinting (ERIC-PCR, REP-PCR, BOX-PCR, and RAPD), REP-PCR showed maximum differentiation among strains within each group. Only Chelatococcus strains showed the ability to solubilize and utilize coal as the sole source of carbon. On the basis of 16S rRNA gene sequence and the ability to utilize different carbon sources, the Chelatococcus strains showed maximum similarity to C. daeguensis. This is the first report showing occurrence of Rhizobium and Chelatococcus strains in an Indian coal bed, and the ability of Chelatococcus isolates to solubilize and utilize coal as a sole source of carbon for their growth.

• The Plant Pathology Journal
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• 제37권5호
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• pp.476-488
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• 2021

Xanthomonas campestris pv. campestris (Xcc) is the causal agent of black rot for cruciferous vegetables worldwide, especially for the cole crops such as cabbage and cauliflower. Due to the lack of resistant cabbage cultivars, black rot has brought about considerable yield losses in recent years in China. Understanding of the pathogen features is a key step for disease prevention, however, the pathogen diversity, population structure, and virulence are largely unknown. In this study, we studied 50 Xcc strains including 39 Xcc isolates collected from cabbage in 20 regions across China, using multilocus sequence genotyping (MLST), repetitive DNA sequence-based PCR (rep-PCR), and pathogenicity tests. For MLST analysis, a total of 12 allelic profiles (AP) were generated, among which the largest AP was AP1 containing 32 strains. Further cluster analysis of rep-PCR divided all strains into 14 DNA groups, with the largest group DNA I comprising of 34 strains, most of which also belonged to AP1. Inoculation tests showed that the representative Xcc strains collected from diverse regions performed differential virulence against three brassica hosts compared with races 1 and 4. Interestingly, these results indicated that AP1/DNA I was not only the main pathotype in China, but also a novel group that differed from the previously reported type races in both genotype and virulence. To our knowledge, this is the first extensive genetic diversity survey for Xcc strains in China, which provides evidence for cabbage resistance breeding and opens the gate for further cabbage-Xcc interaction studies.

• Journal of Microbiology
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• 제44권1호
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• pp.106-112
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• 2006

In this study, the genetic diversities of multi-resistant Salmonella typhimurium (ST) isolates were analyzed via the application of both pulsed field gel electrophoresis (PFGE) and Polymerase chain reaction (PCR) analysis methods, using 6 kinds of primers (REP, ERIC, SERE, BOX, P-1254 and OPB-17). And their discriminative abilities (DA) were also compared in order to determine the most effective and reliable analysis method. 118 S. typhimurium isolates, cultured from diverse animals and human patients in Korea beginning in 1993, were analyzed and subjected to a comparison of Simpson's index of diversity (SID), using both PFGE and PCR methods. PFGE by XbaI enzyme digestion allowed for discrimination into 9 pulsotypes, with high SID values (0.991) on the genomic DNA level. This shows that PFGE is a very discriminative genotypic tool, and also that multiple clones of S. typhimurium isolates had existed in domestic animals and humans in Korea since 1993. However, we could ultimately not to trace the definitive sources or animal reservoirs of specific S. typhimurium isolates examined in this study. Depending on the SID values, the combined method (7 kinds of method) was found to be the most discriminative method, followed by (in order) SERE-PCR, REP-PCR, ERIC-PCR, PFGE & OPB-17 (RAPD), P-1254 (RAPD), and BOX-PCR at the $80\%$ clone cut-off value. This finding suggests that the REP-PCR method (which utilizes 4 primer types) may be an alternative tool to PFGE for the genotyping of S. typhimurium isolates, with comparable cost, time, and labor requirement. The establishment of a highly reliable and discriminatory method for epidemiologic analysis is considered necessary in order for researchers to trace the sources of specific pathogens and, consequently, to control and prevent the spread of epidemic S. typhimurium isolates to humans.

• 대한임상검사과학회지
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• 제36권1호
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• pp.1-6
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• 2004

The enterobacterial repetitive intergenic consensus (ERIC)-PCR is a recently described DNA fingerprinting technique based on amplification of repetitive element distributed in bacteria. We applied of ERIC-PCR to clinical isolates of Vibrio parahaemolyticus and other bacteria associated diarrhea. Twenty isolates of V. parahaemolyticus were used for intragenic genotyping, which were isolated from 2001 to 2002 in Chungbuk National University hospital. For interspecies genotyping, V. vulnificus, V. alginolyticus, V. parahaemolyticus, Escherichia coli, Salmonella and Shigella spp. were used. The genotyping were analyzed by ERIC-PCR. The genotyping of V. parahaemolyticus were grouped two major pattern (A, B) and were subdivided into 10 subtypes (A1, A2, B1-B8) by ERIC-PCR. These method distinctly differentiated bacterial species associated diarrhea. Those results show that ERIC-PCR can be reliable and efficient method for genotyping of V. parahaemolyticus and bacteria associated diarrhea.

• Food Science and Biotechnology
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• 제17권6호
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• pp.1171-1177
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• 2008

This study was undertaken to classify Enterobacter sakazakii isolates from 13 powdered infant formula products, 25 powdered weaning diet products, and 33 weaning diet ingredients on polymerse chain reaction (PCR) methods. The numbers of the isolates from 1 powdered infant formula product, 7 powdered weaning diet products, and 6 weaning diet ingredients were 1, 14, and 8, respectively. The contaminated ingredients were 1 rice powder, 2 millet powders, 2 vegetable powders, and 1 fruit and vegetable premix. PCR with the primer of repetitive extragenic palindromic element (REP-PCR) and random amplification of polymorphic DNA(RAPD) were effective in discriminating among the isolates, but tRNA-PCR and PCR with the primer of l6S-23S internal transcribed spacer (ITS-PCR) were not. Some of E. sakazakii isolates from vegetable powders, fruit and vegetable premix, and millets powders were classified into the clonal groups based on the DNA patterns in the REP-PCR and RAPD analysis. A close genetic relationship among the isolates from some of the powdered weaning diet products and the rice powder was also detected in the cluster analysis based on the DNA patterns in RAPD.

• 식물병연구
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• 제19권4호
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• pp.265-272
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• 2013

전국 7개도 14개 시 군의 주요 고추 재배지에서 시들음 증상을 나타내는 식물체의 땅가 줄기에서 분리 선별된 63개 풋마름병균들의 특성을 조사하였다. 고추에서 분리된 풋마름병균들은 고추(cv. 대왕)와 토마토(cv. 서광)에 강한 병원성을 나타냈다. 모든 병원균들은 배양적, 생리 생화학적 특성 및 특이 PCR 검출에 의하여 Ralstonia solanacearum으로 동정되었다. 고추에서 분리된 63개 풋마름병균들은 race 1 계통으로 2가지 biovar로 구분되었는데, biovar 3은 17개 균주로 27%였고 biovar 4는 46개의 균주로 73%였으며, biovar 4의 생리형이 우점계통이었다. Rep-PCR에 의한 국내 고추 풋마름병균들의 유전적 다양성을 확인한 결과, 70%의 유사성을 기준으로 12개의 group으로 구분되었다. 이러한 결과들은 국내 고추 풋마름병에 대한 방제와 저항성 품종 육성에 중요한 기초 자료를 제공할 것이다.

• 대한임상검사과학회지
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• 제48권4호
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• pp.327-334
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• 2016

최근 사람과 가축에 항균제의 과도한 사용으로 감염병을 일으키는 병원성 세균들의 항균제 내성이 증가하고 있다. 본 연구에서는 PCR과 염기서열분석법을 이용하여 충청지역 일개의 대학병원에 의뢰된 임상검체와 같은 지역에서 사육된 닭으로부터 분리된 P. mirabilis 균주를 대상으로 16S ribosomal RNA methyltransferase(RMTase) 유전자와 integron을 조사하였다. 또한 Repetitive extragenic palindromic sequence-based PCR (REP-PCR)을 이용하여 P. mirabilis 균주들의 역학적 연관성 조사하였다. 총 38균주의 P. mirabilis 중에서 임상검체로부터 분리된 7균주 (18.4%)만이 RMTases 유전자를 가지고 있었는데 이들은 모두 amikacin, tobramycin, 및 gentamicin에 내성을 나타냈다. 또한 대상균주 중 23균주(60.5%)가 class 1 integron을 가지고 있는 것으로 나타났으며 class 2 및 class 3 integron은 검출되지 않았다. 본 연구에서 확인된 integrons에는 aminoglycoside 내성유전자(aadA2, aadA5, aadA7, 및 aacCA5), ${\beta}$-lactmam 내성유전자($bla_{PSE}$), erythromycin 내성유전자(ereA), lincosamides 내성유전자(linF), 및 trimethoprim 내성유전자(dfrA12, dfrA17 및 dfrA32)등이 유전자 카세트로 포함되어 있었다. 본 연구결과 RMTase 유전자는 임상검체로부터 분리된 P. mirabilis 균주에만 확산되어 있었던 반면 class 1 integrons는 임상검체와 닭으로부터 분리된 P. mirabilis 균주에 광범위하게 확산되어 있음을 확인할 수 있었다. 게다가 닭으로부터 분리된 균주 중에는 동일한 REP-PCR 밴드패턴을 보인 균주들이 있었는데 이는 닭들 사이에서 P. mirabilis 균주가 수평확산 되었음을 의미한다. P. mirabilis 균주에서 항균제 내성유전자의 확산을 막기 위해서는 내성유전자 지속적인 모니터링과 감시가 필요할 것으로 사료된다.

• 대한임상검사과학회지
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• 제48권2호
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• pp.94-101
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• 2016

Acinetobacter species는 중요한 기회감염균으로 빈번하게 원내감염을 일으킨다. 뿐만 아니라 다제내성을 보이는 경우가 많아 치료를 위한 항균제 선택이 매우 제한적이다. 본 연구에서는 Acinetobacter species 68균주를 대상으로 하여 다양한 carbapenemase 유전자를 조사했다. 디스크확산법으로 항균제 감수성 양상을 조사했으며 다중 중합효소연쇄반응을 통해 carbapenemase 유전자를 포함하고 있는 균주를 선별하였고 PCR과 염기서열분석을 통해 최종적으로 carbapenemase 유전형을 확인했다. 한편 REP-PCR 방법으로 균주간의 clonality를 분석했다. 본 연구에서 A. baumannii 균주는 분석된 모든 항균제에 대해 높은 내성을 나타냈으나 non-A. baumannii 균주는 분석된 항균제 중 aztreonam과 cefotaxime을 제외한 항균제에 모두 감수성을 보였다. A. baumannii 51 균주는 $bla_{OXA-51}$ 유전자를 가지고 있었으며 그 중 37(72.5%)균주는 $bla_{OXA-23}$ 유전자도 동시에 가지고 있었다. 본 연구에서 39 균주의 다제내성 A. baumannii 가 분리되었는데 그 중 37균주가 $bla_{OXA-23}$ 유전자를 가지고 있었다. $bla_{OXA-23}$ 유전자를 포함하고 있는 균주들은 I (n=22) 형 또는 II (n=15) 형의 REP-PCR band 패턴을 보였는데 이는 대전에 위치한 일개의 대학병원에 $bla_{OXA-23}$ 유전자를 포함하고 있는 다제내성 균주들이 수평 확산 되어 있음을 의미한다. 다제내성 A. baumannii 균주에 의한 감염 및 집락화를 막기 위해서는 지속적으로 내성을 유발하는 인자를 조사하고 MDR균주의 출현 및 확산을 감시할 필요가 있을 것으로 사료된다.

• 식물병연구
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• 제15권2호
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• pp.77-82
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• 2009

최근 혹병이 심하게 발생한 충북지방의 포도나무의 뿌리로부터 선택배지를 이용하여 Agrobacterium속 세균을 분리하고 동정하였다. 분리균의 지방산 분석을 통한 MIDI에 의한 동정, 16S rDNA 염기서열, 생화학적 특성, 종 특이적 primer을 이용한 PCR 결과로 13개 분리균은 모두 A. tumefaciens로 동정 되었으며, 포도나무 혹병균인 A. vitis는 분리되지 않았다. 모든 분리균은 토마토와 포도나무의 줄기와 뿌리에 병원성을 나타내지 않았다. Rep-PCR 결과 분리균은 A. tumefaciens와 일부 유사성이 있지만 전체적으로 병원균인 A. tumefaciens와 A. vitis와는 유사성이 낮았다. 이상의 결과는 충북 일부 지방의 MBA와 거봉에서 발생한 혹병을 일으키는 병원균은 토양이나 뿌리로부터 전반되지 않고 묘목을 통해서 전반되었을 가능성을 시사하고 있다.