• Title/Summary/Keyword: Prevotella intermedia ATCC 49046

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Strain-specific PCR Primers for the Detection of Prevotella intermedia ATCC 49046

  • Kim, Min-Jung;Min, Jeong-Bum;Lim, Sun-A;Kook, Joong-Ki
    • International Journal of Oral Biology
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    • v.36 no.2
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    • pp.79-82
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    • 2011
  • The aim of this study was to develop Prevotella intermedia ATCC 49046-specific PCR primers designed based on the nucleotide sequence of a DNA probe Pig28. The strainspecificity of the PCR primers, Pig28-F1/Pig28-R1, was confirmed with 9 strains of P. intermedia and 25 strains (15 species) of Prevotella species. The detection limit of the PCR primers was 2 pg of the purified genomic DNA of P. intermedia ATCC 49046. These PCR primers were found to be useful for identifying P. intermedia ATCC 49046, particularly for determining the authenticity of the strain.

Development of prevotella intermedia ATCC 49046 Strain-Specific PCR Primer Based on a Pig6 DNA Probe (Pig6 DNA probe를 기반으로 하는 Prevotella intermedia ATCC 49046 균주-특이 PCR primer 개발)

  • Jeong Seung-U;Yoo So-Young;Kang Sook-Jin;Kim Mi-Kwang;Jang Hyun-Seon;Lee Kwang-Yong;Kim Byung-Ok;Kook Joong-Ki
    • Korean Journal of Microbiology
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    • v.42 no.2
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    • pp.89-94
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    • 2006
  • The purpose of this study is to develop the strain-specific PCR primers for the identification of prevotella inter-media ATCC 49046 which is frequently used in the pathogenesis studies of periodontitis. The Hind III-digested genomic DNA of P. intermedia ATCC 49046 were cloned by random cloning method. The specificity of cloned DNA fragments were determined by Southern blot analysis. The nucleotide sequence of cloned DNA probes was determined by chain termination method. The PCR primers were designed based on the nucleotide sequence of cloned DNA fragment. The data showed that Pig6 DNA probe were hybridized with the genomic DNA from P. intermedia strains (ATCC $25611^T$ and 49046) isolated from the Westerns, not the strains isolated from Koreans. The Pig6 DNA probe were consisted of 813 bp. Pig6-F3 and Pig6-R3 primers, designed base on the nucleotide Sequences Of Pig6 DNA Probe, were 3150 specific to the only both P. intermedia ATCC $25611^T$ and P. intermedia ATCC 49046. In the other hand, Pig6-60F and Pig6-770R primers were specific to the only P. intermedia ATCC 49046. The two PCR primer sets could detect as little as 4 pg of chromosomal DNA of P. intermedia. These results indicate that Pig6-60F and Pig6-770R primers have proven useful for the identification of P. intermedia ATCC 49046, especially with regard to the maintenance of the strain.

Identification of Prevotella intermedia ATCC 25611 Using Pi29-L DNA Probe. (Pi29-L DNA 프로브를 이용한 Prevotella intermedia ATCC 25611의 동정)

  • 국중기;백동헌
    • Microbiology and Biotechnology Letters
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    • v.31 no.2
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    • pp.205-209
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    • 2003
  • Recently, we introduced a new method for rapid screening of bacterial species- or subspecies-specific DNA probes, named “inverted dot blot hybridization screening method”. We then applied this method to develop species- or strain- specific DNA probes for Prevotella intermedia and Prevotella nigrescens. In those studies, among 96 candidate DNA probes which were screened by the new method, 5 probes were confirmed as being putatively strain-specific : 3 probes for P. nigrescens 9336 (ATCC 33563), one for each p. intermedia ATCC 25611 and one for P. nigrescens G8-9K-3 (ATCC 49046). In the present study, we evaluated by Southern blot analysis a DNA probe Pi29-L, one of the 96 candidate probes described above, whether it is specific for the strain ATCC 25611 off. intermedia. Our data show that the probe Pi29-L is potentially P. intermedia ATCC 25611-specific, which can be useful for the detection and identification of the strain, particularly in maintenance of the strain.

EFFECT OF POLYPHOSPHATE ON THE GROWTH OF ORAL BACTERIUM, PREVOTELLA INTERMEDIA (구강세균 Prevotella intermedia의 성장에 따른 polyphosphate의 영향에 관한 연구)

  • Kong, Hee-Joung;Choi, Ho-Young;Min, Byung-Soon;Part, Sang-Jin;Lee, Jin-Yong;Choi, Gi-Woon
    • Restorative Dentistry and Endodontics
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    • v.23 no.2
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    • pp.550-560
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    • 1998
  • Prevotella intermedia has been known as one of the important bacterial species involved in the endodontic infections and various periodontal diseases. Polyphosphate has been widely used to prevent decomposition of food and known to have an inhibitory effect on the growth of gram positive bacteria. The purpose of this study was to evaluate the effect of poly phosphate on the growth of Prevotella intermedia, a gram negative bacterium. Prevotella intermedia G8GK3(ATCC 49046) was grown in the presence of polyphosphates with different chain lengths. Inhibitory effect of each polyphosphate, which was added at the beginning or at the early exponential growth phase of Prevotella intermedia, was determined by measuring optical density of the bacterial cells at 540nm, viable cells and lysis of Prevotella intermedia. The results from this study were as follows : 1. Poly phosphate inhibited the growth of Prevotella intermedia. 2. The minimum inhibitory concentration(MIC) of poly phosphate appeared to be 0.05%. 3. Polyphosphates with chain lengths of 5 and 65 demonstrated the greatest inhibitory effect on the growth of Prevotella intermedia. 4. Polyphosphate was bactericidal to Prevotella intermedia, demonstrating the growth inhibition of the bacterium. 5. Polyphosphate induced lysis of Prevotella intermedia. The overall results suggest that polyphosphate has a bactericidal effect on Prevotella intermedia, causing the lysis of the bacterium.

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Antimicrobial Activity of Oleanolic Acid, Ursolic Acid, and Sophoraflavanone G against Periodontopathogens

  • Park, Soon-Nang;Kook, Joong-Ki
    • International Journal of Oral Biology
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    • v.38 no.4
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    • pp.149-154
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    • 2013
  • In general, oleanolic acid (OA) and ursolic acid (UA) have antimicrobial effect against Gram-positive bacteria but not Gram-negative bacteria whereas sophoraflavanone G has antimicrobial activity against both bacterial types. However, the antimicrobial effects of OA, UA, and sophoraflavanone G against periodontopathogens have not been studied to any great extent. The aim of this study was to investigate antimicrobial effect of OA, UA, and sophoraflavanone G against 15 strains (5 species) of oral Gram-negative bacteria, which are the major causative bacteria of periodontal disease. The antimicrobial activity was evaluated by minimal inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) determinations. OA and UA showed antimicrobial effects against all of the Porphyromonas gingivalis strains tested and also Prevotella intermedia ATCC $25611^T$. Interestingly, P. intermedia ATCC 49046 showed greater resistance to OA and UA than P. intermedia ATCC $25611^T$. In contrast, sophoraflavanone G had antimicrobial activity against all strains, with MIC and MBC values below $32{\mu}g/ml$, except Aggregatibacter actinomycetemcomitans. These results indicate that sophoraflavanone G may have potential for use in future oral hygiene products such as dentifrices and gargling solution to prevent periodontitis.