• Title, Summary, Keyword: F. nucleatum

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Identification of Fusobacterium nucleatum isolated from Korean by F. nucleatum subspecies-specific DNA probes (Dot blot hybridization법을 이용한 Fusobacterium nucleatum 아종-특이 DNA 프로브의 특이성 평가)

  • Kim, Hwa-Sook;Kook, Joong-Ki
    • Journal of Korean society of Dental Hygiene
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    • v.6 no.4
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    • pp.311-324
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    • 2006
  • The purpose of this investigation was to evaluate of the specificity of Fusobacterium nucleatum subspecies-specific DNA probes using dot blot hybridization. To confirm whether the clinical isolates were F. nucleatum or not, 16S rDNA of them were cloned and sequenced. The sequencing data were used in homology search with database of GenBank. When the homology was above 98% compared with the nucleotide sequence of a certain bacteria, it was judged as the same species with the bacteria. 23 strains of F. nucleatum were isolates from subgingival plaque of periodontitis patient. The clinical isolates of F. nucleatum were classified into 10 groups using phylogenetic analysis of 16S rDNA sequence. F. nucleatum subspecies nucleatum-specific DNA probe Fu4(1.3 kb) reacted with genomic DNAs from 8 type strains of F. nucleatum and it reacted strongly with those from 8 clinical isolates. The Fp4(0.8 kb) reacted with F. nucleatum subsp. polymorphum ATCC 10953 and one clinical isolates. Fv35(1.9 kb) and Fs17(8.2 kb) probes reacted with genomic DNAs from F. nucleatum subsp. vincentii ATCC 49256 and F. nucleatum subsp. fusiform ATCC 51190, respectively. Our results showed that it is not enough to evaluate the specificity of F. nucleatum subspecies-specific DNA probes with only dot blot hybridization. Therefore, Southern blot analysis will be necessary to confirm the specificity of F. nucleatum subspecies-specific DNA probes.

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Complete genome sequence of Fusobacterium nucleatum KCOM 1323 isolated from a human subgingival plaque of periodontitis lesion (사람 치주질환병소의 치은연하지면세균막에서 분리된 Fusobacterium nucleatum KCOM 1323의 유전체 염기서열 해독)

  • Park, Soon-Nang;Lim, Yun Kyong;Shin, Ja Young;Roh, Hanseong;Kook, Joong-Ki
    • Korean Journal of Microbiology
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    • v.53 no.3
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    • pp.219-221
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    • 2017
  • Fusobacterium nucleatum is a Gram-negative, obligately anaerobic and rod- or filament-shaped bacterium. F. nucleatum is part of oral microflora and is a causative agent of periodontitis as well as is associated with a wide spectrum of systemic diseases of human. F. nucleatum KCOM 1323 (= ChDC F317) was isolated from a human subgingival plaque of periodontitis lesion. Here, we present the complete genome sequence of F. nucleatum KCOM 1323.

Development of Strain-specific PCR Primers Based on a DNA Probe Fu12 for the Identification of Fusobacterium nucleatum subsp. nucleatum ATCC $25586^T$

  • Kim Hwa-Sook;Song Soo Keun;Yoo So Young;Jin Dong Chun;Shin Hwan Seon;Lim Chae Kwang;Kim Myong Soo;Kim Jin-Soo;Choe Son-Jin;Kook Joong-Ki
    • Journal of Microbiology
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    • v.43 no.4
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    • pp.331-336
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    • 2005
  • The objective of this study was to assess the strain-specificity of a DNA probe, Fu12, for Fusobacterium nucleatum subsp. nucleatum ATCC $25586^T$ (F. nucleatum ATCC $25586^T$), and to develop sets of strain-specific polymerase chain reaction (PCR) primers. Strain-specificity was tested against 16 strains of F. nucleatum and 3 strains of distinct Fusobacterium species. Southern blot hybridization revealed that the Fu12 reacted exclusively with the HindIII-digested genomic DNA of F. nucleatum ATCC $25586^T$. The results of PCR revealed that three pairs of PCR primers, based on the nucleotide sequence of Fu12, generated the strain-specific amplicons from F. nucleatum ATCC $25586^T$. These results suggest that the DNA probe Fu12 and the three pairs of PCR primers could be useful in the identification of F. nucleatum ATCC $25586^T$, especially with regard to the determination of the authenticity of the strain.

Prognostic Impact of Fusobacterium nucleatum Depends on Combined Tumor Location and Microsatellite Instability Status in Stage II/III Colorectal Cancers Treated with Adjuvant Chemotherapy

  • Oh, Hyeon Jeong;Kim, Jung Ho;Bae, Jeong Mo;Kim, Hyun Jung;Cho, Nam-Yun;Kang, Gyeong Hoon
    • Journal of Pathology and Translational Medicine
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    • v.53 no.1
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    • pp.40-49
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    • 2019
  • Background: This study aimed to investigate the prognostic impact of intratumoral Fusobacterium nucleatum in colorectal cancer (CRC) treated with adjuvant chemotherapy. Methods: F. nucleatum DNA was quantitatively measured in a total of 593 CRC tissues retrospectively collected from surgically resected specimens of stage III or high-risk stage II CRC patients who had received curative surgery and subsequent oxaliplatin-based adjuvant chemotherapy (either FOLFOX or CAPOX). Each case was classified into one of the three categories: F. nucleatum-high, -low, or -negative. Results: No significant differences in survival were observed between the F. nucleatum-high and -low/negative groups in the 593 CRCs (p=.671). Subgroup analyses according to tumor location demonstrated that disease-free survival was significantly better in F. nucleatum-high than in -low/negative patients with non-sigmoid colon cancer (including cecal, ascending, transverse, and descending colon cancers; n=219; log-rank p=.026). In multivariate analysis, F. nucleatum was determined to be an independent prognostic factor in non-sigmoid colon cancers (hazard ratio, 0.42; 95% confidence interval, 0.18 to 0.97; p=.043). Furthermore, the favorable prognostic effect of F. nucleatum-high was observed only in a non-microsatellite instability-high (non-MSI-high) subset of non-sigmoid colon cancers (log-rank p=0.014), but not in a MSI-high subset (log-rank p=0.844), suggesting that the combined status of tumor location and MSI may be a critical factor for different prognostic impacts of F. nucleatum in CRCs treated with adjuvant chemotherapy. Conclusions: Intratumoral F. nucleatum load is a potential prognostic factor in a non-MSI-high/non-sigmoid/non-rectal cancer subset of stage II/III CRCs treated with oxaliplatin-based adjuvant chemotherapy.

Cellular and Humoral Immune Responses to Sequential Periodontopathic Bacterial Immunization in Animal Model (상이한 치주병원균의 연속적 인공면역에 대한 세포성 및 체액성 면역반응에 대한 동물실험적 연구)

  • Jeon, Soo-Kyung;Kim, Sung-Jo;Choi, Jeom-Il
    • Journal of Periodontal and Implant Science
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    • v.30 no.3
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    • pp.687-700
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    • 2000
  • Antigen-specific T cell clones were obtained from mice immunized with Fusobacterium nucleatum ATCC 10953(F .nucleatum) and/or Porphyromonas gingi valis 381(P. gingivalis). 10 Balb/c mice per group were immunized with F. nucleatum followed by P. gingivalis, or with P. gingivalis alone by intraperitoneal injection of viable microorganisms. Spleen T cells were isolated and stimulated in vitro with viable P. gingivalis cells to establish P. gingivalisspecific T cell clones. T cell phenotypes and cytokine profiles were determined along with T cell responsiveness to F .nucleatum or P. gingivalis. Serum IgG antibody titers to F. nucleatum or P. gingivalis were also determined by ELISA. All the T cell clones derived from mice immunized with F. nucleatum followed by P. gingivalis demonstrated Th2 subsets, while those from mice immunized with P. gingivalis alone demonstrated Th1 subsets based on the flow cytometric analysis and cytokine profiles, All T cells clones from both groups were cross-reactive to both P. gingivalis and F. nucleatum antigens. Phenotypes of T cell clones were all positive for CD4. Mean post-immune serum IgG antibody levels to F. nucleatum or P . gingivalis were significantly higher than the pre-immune levels(p <0.01, respectively). There were no significant differences in the antibody titers between the two groups. It was concluded that P. gingivalis-specific T cells initially primed by cross-reactive F. nucleatum antigens were polarized to Th2 subsets, while T cells stimulated with P. gingivalis alone maintained the profile of Th1 subset.

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IDENTIFICATION OF FUSOBACTERIUM NUCLEATUM AND FUSOBACTERIUM NECROPHORUM USING POLYMERASE CHAIN REACTION(PCR) (중합효소연쇄반응(Polymerase Chain Reaction)을 이용한 Fusobacterium nucleatum 및 Fusobacterium necrophorum의 동점에 관한 연구)

  • Kang, Chang-Woo;Park, Dong-Sung;Yoon, Soo-Han
    • Restorative Dentistry and Endodontics
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    • v.24 no.1
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    • pp.40-48
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    • 1999
  • This study was designed to examine the specificities of the designed primers for F. nucleatum and F. necrophorum and to compare the PCR results using clinical samples with those of the anaerobic culture method. F. nucleatum and F. necrophorum spp. are frequently isolated in infected root canals, and they are related to periapical diseases. F. nucleatum(VPI 10197) and F. necrophorum(ATCC 25286) were used as references for PCR reaction, and thirty five teeth with one canal and periapical lesion were used. The samples were cultured anaerobically and identified using Rapid ID 32A(BioMerieux Vitek, Inc., France) as biochemical battery. In the GenBank database, species-specific PCR primers(nuc1/nuc2 primers for F. nucleatum and nec1/nec2 primers for F. necrophorum) were designed from the 16S ribosomal DNA(rDNA) sequences of F. nucleatum(accession number M58683) and F. necrophorum(accession number AF044948). PCR procedures of F. nucleatum(VPI 10197) and F. necrophorum (ATCC 25286) were simulated on a computer software. Amplify(v.1.2${\beta}$ for Macintosh). 820 bps and 817 bps of nucleotides were expected, respectively. Using extracted DNAs with QiaAmp tissue kit(Qiagen co., Germany), PCR was done. The results were as follows : 1. The nuc1/nuc2 primers produced an amplicon of 820 bps and the nec1/nec2 primers produced an amplicon of 817 bps. 2. The nuc1/nuc2 primers and the nec1/nec2 primers were specific and did not react with species other than the designated ones(i.e. nuc1/nuc2 primers did not produce amplicons for F. necrophorum, and vice versa.). And the PCR products of Porphyromonas endodontalis(ATCC 35406), Porphyromonas gingivalis(ATCC 33277), Prevotella intermedia(ATCC 25611), and Prevotella nigrescens(ATCC 33563), frequently isolated in infected root canals and periapical lesions, were not amplified by the primers specific for Fusobacterium nucleatum and Fusobacterium necrophorum. 3. This method utilizing PCR could detect F. nucleatum and F. necrophorum in clinical samples, while anaerobic culture method could detect neither.

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Isolation of Fusobacterium nucleatum from subgingival plaque in Korean (한국인의 치은연하 치태에서 Fusobacterium nucleatum의 분리)

  • Jang, Hyun-Seon;Kim, Seo-Hoon;Kim, Hwa-Sook;Kook, Joong-Ki;Kim, Mi-Kwang;Yoo, So-Young;Kim, Byung-Ock
    • Journal of Periodontal and Implant Science
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    • v.33 no.2
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    • pp.149-158
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    • 2003
  • The purpose of this study was to isolate and characterize the Fusohacrerium nucleatum (F. nucleatum) from subgingival plaque in Korean periodontitis patients. The subgingival plaque samples of periodontitis patient were collected with sterilized paper point. The paper point was put into reduced transfer medium and then immediately transferred to laboratory. The subgingival samples were diluted by 10,000 folds and plated on F. nucleatum-selective media agar plate. The plates were incubated at 37$^{\circ}C$ in an anaerobic chamber for 3 days. The violet-colored colonies were selected and subjected to further verification whether those are F. nucleatum or not. For further confirmation, 16S rRNA genes (rDNA) were cloned from each of bacterial clones and determined sequence of 16S rDNA. In this study, we found 17 distinct clinical isolates of F. nucleatum from subgingival plaque. The clinical isolates will be a useful in various studies in periodontology.

Antibacterial Effect on Oral Normal flora of Phytoncide from Chamaecyparis Obtusa (구강 상주균에 대한 편백 피톤치드의 항균효과)

  • Auh, Q-Schick;Hong, Jung-Pyo;Chun, Yang-Hyun
    • Journal of Oral Medicine and Pain
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    • v.34 no.4
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    • pp.353-362
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    • 2009
  • The present study was performed to observe the effect of phytoncide on oral normal microflora and the inhibitory effect of the surviving resident oral bacteria on F. nucleatum. In this study, saliva from each of 20 healthy subjects was treated with 1% phytoncide from Japanese Hinoki (Chamaecyparis obtusa Sieb. et Zucc.). The surviving salivary bacterium were isolated on blood agar plates and identified by 16S rDNA sequencing. In order to select inhibitory isolates against F. nucleatum, the isolates from the phytoncide-treated saliva were cultured with F. nucleatum. The results are as follows: 1. Among the 200 surviving resident oral bacterium, 70(35.0%) bacterium inhibit the growth of F. nucleatum on blood agar plates. 2. Among the 70 bacterium which inhibit F. nucleatum, Streptococcus salivarius was 41.3%(45/109), Streptococcus sanguinis was 28%.(7/25), Streptococcus mitis was 20%(3/15), Streptococcus parasanguinis was 33.3%(3/9), Streptococcus Alactolyticus was 100%(8/8), Streptococcus vestibularis was 28.6%(2/7) and Streptococcus sp. was 50%(2/4). Taken together, among the surviving resident oral bacterium, Streptococcus salivarius, Streptococcus sanguinis, Streptococcus mitis were mainly observed to inhibit F. nucleatum. and they may exert an additional inhibitory activity against the periodontopathic bacterium. Therefore, phytoncide can be used to prevent and cease the progress of periodontal disease, halitosis. Thus it is expected to promote oral health.

Comparison between Bacterial Culture Method and Multiplex PCR for Identification of Fusobacterium nucleatum and Actinobacillus actinomycetemcomitans from the Dental Plaques (치면세균막내의 Fusobacterium nucleatum과 Actinobacillus actinomycetemcomitans의 동정을 위한 세균배양법 및 Multiplex PCR법의 비교)

  • Kim, Hwa-Sook;Lim, Sun-A
    • Journal of dental hygiene science
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    • v.9 no.2
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    • pp.249-255
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    • 2009
  • This study was carried out for the purpose of comparing bacterial culture method, single PCR, and multiplex PCR for identification of F. nucleatum and A. actinomycetemcomitans in subgingival plaque of adult periodontitis. Targeting 20 patients with adult periodontitis, the subgingival plaque was collected in teeth, respectively, for #16, #36, #44. A bacillus was cultivated by painting it over the solid selective media of F. nucleatum and A. actinomycetemcomitans. Bacterial species were detected in 0 tooth with 12 pieces, respectively. Through single PCR and multiplex PCR, the positive reaction was indicated in 43 teeth with 45 pieces, respectively, as for F. nucleatum, and in 1 tooth with 4 pieces, respectively, as for A. actinomycetemcomitans. In the comparative analysis between bacterial identification methods. F. nucleatum showed the more statistically significant difference(p=0.0(0) in comparison between single PCR and multiplex PCR. Even A. actinomycetemcomitans was indicated significantly(p=0.067) in a case that is based on 0.1 in significant level in the comparison between single PCR and multiplex PCR. In conclusion, as a result of comparing the bacterial identification methods, the detection frequency was indicated to be higher in PCR than in bacterial culture method. Single PCR and multiplex PCR showed the mutually similar detection frequency. Accordingly, given thinking of economic efficiency, quickness, and reduction in labor force, it is thought to be more efficient method to use single PCR as the bacterial identification method.

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