Development of quantitative real-time PCR primers for detecting 42 oral bacterial species

Park, Soon-Nang; Lim, Yun Kyong; Kook, Joong-Ki

doi:10.1007/s00203-013-0896-4

Cited by 35 publications

(21 citation statements)

References 31 publications

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“…The gene coding for the RNA polymerase β subunit, rpoB , has been established as a useful taxonomic marker gene in genotypic analyses of Fn and is predicted to have a greater resolving power than those for 16S genes . FISH results in the present study were further validated in 20 samples by Fn ‐specific PCR primers designed to target rpoB as a previous study reported …”

Section: Methodssupporting

confidence: 62%

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Invasive Fusobacterium nucleatum may play a role in the carcinogenesis of proximal colon cancer through the serrated neoplasia pathway

Chen

et al. 2016

Intl Journal of Cancer

133

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The prevalence of invasive Fusobacterium nucleatum (Fn) within the serrated neoplasia pathway of the proximal colon has seldom been investigated. We examined the invasive Fn and bacterial biofilms in 35 proximal hyperplastic polyps (HPs), 33 sessile serrated adenomas (SSAs), 48 proximal colorectal cancers (CRCs) and 10 matched metastatic lymph nodes using 16S rRNA fluorescence in situ hybridization (FISH). Samples of normal mucosa, traditional adenomas (TAs), distal HPs, distal CRCs and matched lymph nodes with or without metastases were used as controls. The prevalence of invasive Fn within proximal HPs (65.7%) and SSAs (78.8%) were significantly higher than that of proximal TAs (28.9%) and distal TAs (24.4%; p < 0.05). Invasive Fn was detected in markedly more proximal CRCs (89.6%) than in distal CRCs (42.2%; p < 0.05). Moreover, invasive Fn was detected in a significantly higher proportion of matched metastatic lymph nodes (100%) than that within nonmetastatic lymph nodes (40.0%; p < 0.001). Bacterial biofilms were found on 52.1% of proximal CRCs, 55.6% of distal CRCs and 48.5% of SSAs. Biofilms were positive for Fn in 47.9% of proximal CRCs, 48.9% of distal CRCs and 27.3% of SSAs. However, the presence of Fn in biofilms was not related to invasive Fn within colorectal tissues (p 5 0.415). Invasive Fn may play a role in the carcinogenesis of proximal colon developing via the serrated neoplasia pathway, but might have a less important role in the TA-carcinoma sequence. Bacterial biofilms may not contribute to the invasion of Fn into tumor tissues.The human gut is populated by hundreds of different bacterial species, some of which contribute toward the pathogenesis of a range of human diseases.1 Recently, Fusobacterium nucleatum (Fn) has been recognized as an opportunistic pathogen implicated in inflammatory diseases of the gut, including appendicitis and inflammatory bowel diseases. 2,3Current microbiome surveys of human tumors revealed an enrichment of Fn in colorectal cancer (CRC). 4-8 Moreover,accumulating evidence suggest that Fn may play a role in modulating CRC development. 4,9Many of these observations, however, were studies of the traditional adenoma (TA)-carcinoma sequence. In recent years, it's believed that up to 20% of all CRCs arise through an alternative, serrated neoplasia pathway.

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Section: Methodssupporting

confidence: 62%

“…25 FISH results in the present study were further validated in 20 samples by Fn-specific PCR primers designed to target rpoB as a previous study reported. 26…”

Section: Validation Of Fish Results By Conventional Pcrmentioning

confidence: 99%

Invasive Fusobacterium nucleatum may play a role in the carcinogenesis of proximal colon cancer through the serrated neoplasia pathway

Chen

et al. 2016

Intl Journal of Cancer

133

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“…14, 15 The PCR primers of Pi and Pg were designed based on the 16S rRNA gene (Table 2). A conserved sequence in the 16S rRNA was selected to count the total bacteria.…”

Section: Methodsmentioning

confidence: 99%

Changes in salivary periodontal pathogens after orthodontic treatment: An in vivo prospective study

Kim

Jung

Cho

et al. 2015

The Angle Orthodontist

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Objective: To analyze the initial changes in salivary levels of periodontal pathogens after orthodontic treatment with fixed appliances. Materials and Methods: The subjects consisted of 54 adult patients. The Simplified Oral Hygiene Index, Plaque Index, and Gingival Index were measured as periodontal parameters. Both the plaque and gingival indexes were obtained from the central and lateral incisors and first molars of both arches. Whole saliva and periodontal parameters were obtained at the following four time points: immediately before debonding (T1), 1 week after debonding (T2), 5 weeks after debonding (T3), and 13 weeks after debonding (T4). Repeated measures analysis of variance was used to determine salivary bacterial levels and periodontal parameters among the four time points after quantifying salivary levels of Aggregatibacter actinomycetemcomitans (Aa), Fusobacterium nucleatum (Fn), Porphyromonas gingivalis (Pg), Prevotella intermedia (Pi), Tannerella forsythia (Tf), and total bacteria using the real-time polymerase chain reaction. Results: All periodontal parameters were significantly decreased immediately after debonding (T2). The salivary levels of total bacteria and Pg were decreased at T3, while Pi and Tf levels were decreased at T4. However, the amount of Aa and Fn remained at similar levels in saliva during the experimental period. Interestingly, Aa and Fn were present in saliva at higher levels than were Pg, Pi, and Tf. Conclusion: The higher salivary levels of Aa and Fn after debonding suggests that the risk of periodontal problems cannot be completely eliminated by the removal of fixed orthodontic appliances during the initial retention period, despite improved oral hygiene. (Angle Orthod. 2016;86:998-1003.)

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“…DNA was extracted from positively identified isolates using TIANamp Bacteria DNA Kit. Aliquots were used as template DNA for a PCR with primers and conditions as described by Park et al [ 38 ]. An amplicon size of 171 bp confirmed that the isolate belonged to Fn .…”

Section: Methodsmentioning

confidence: 99%

Invasive Fusobacterium nucleatum activates beta-catenin signaling in colorectal cancer via a TLR4/P-PAK1 cascade

et al. 2017

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The underlying mechanism of Fusobacterium nucleatum (Fn) in the carcinogenesis of colorectal cancer (CRC) is poorly understood. Here, we examined Fn abundance in CRC tissues, as well as β-catenin, TLR4 and PAK1 protein abundance in Fn positive and Fn negative CRCs. Furthermore, we isolated a strain of Fn (F01) from a CRC tissue and examined whether Fn (F01) infection of colon cancer cells activated β-catenin signaling via the TLR4/P-PAK1/P-β-catenin S675 cascade. Invasive Fn was abundant in 62.2% of CRC tissues. TLR4, PAK1 and nuclear β-catenin proteins were more abundant within Fn-positive over Fn-negative CRCs (P < 0.05). Fn and its lipopolysaccharide induced a significant increase in TLR4/P-PAK1/P-β-catenin S675/C-myc/CyclinD1 protein abundance, as well as in the nuclear translocation of β-catenin. Furthermore, inhibition of TLR4 or PAK1 prior to challenge with Fn significantly decreased protein abundance of P-β-catenin S675, C-myc and Cyclin D1, as well as nuclear β-catenin accumulation. Inhibition of TLR4 significantly decreased P-PAK1 protein abundance, and for the first time, we observed an interaction between TLR4 and P-PAK1 using immunoprecipitation. Our data suggest that invasive Fn activates β-catenin signaling via a TLR4/P-PAK1/P-β-catenin S675 cascade in CRC. Furthermore, TLR4 and PAK1 could be potential pharmaceutical targets for the treatment of Fn-related CRCs.

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Development of quantitative real-time PCR primers for detecting 42 oral bacterial species

Cited by 35 publications

References 31 publications

Invasive Fusobacterium nucleatum may play a role in the carcinogenesis of proximal colon cancer through the serrated neoplasia pathway

Invasive Fusobacterium nucleatum may play a role in the carcinogenesis of proximal colon cancer through the serrated neoplasia pathway

Changes in salivary periodontal pathogens after orthodontic treatment: An in vivo prospective study

Invasive Fusobacterium nucleatum activates beta-catenin signaling in colorectal cancer via a TLR4/P-PAK1 cascade

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