Rapid detection of nusG and fadA in Fusobacterium nucleatum by loop-mediated isothermal amplification

Huang, Simo; Yang, Zhan; Zou, Dayang; Dong, Derong; Liu, Anheng; Liu, Wei; Huang, Liuyu

doi:10.1099/jmm.0.000300

Cited by 15 publications

(19 citation statements)

References 27 publications

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“…For S. gallolyticus, the rpoB gene was also sequenced. The presence of the fadA gene in F. nucleatum was determined using the conventional PCR method in accordance with a previous study 31 , and the 211 bp size of the amplicon was confirmed by sequencing.…”

Section: Methodsmentioning

confidence: 79%

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Fusobacterium nucleatum in biopsied tissues from colorectal cancer patients and alcohol consumption in Korea

Kim

Lee

Choi

et al. 2020

Sci Rep

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The roles of individual bacteria and their relationship in the development of colorectal cancer (CRC) remain unclear. We aimed to determine the prevalence of CRC-associated bacteria using quantitative real-time PCR (qPCR) or 16S rRNA analysis and the statistical correlations of patient demographics and clinical characteristics comprising alcohol consumption with CRC-associated bacteria. We determined the prevalence of five CRC-associated bacterial species in 38 CRC patients (39 samples) and 21 normal individuals using qPCR, and the relative abundance of bacterial taxa in the gut microbiome was assessed using 16S rRNA analysis. Fusobacterium nucleatum was the only bacterium that was significantly (P < 0.0001) more prevalent in the cancer tissue (82.1%) than in the normal tissue (0%) by qPCR. 16S rRNA analysis showed a significant correlation between six operational taxonomic units (OTUs), namely, the genera Fusobacterium, Peptostreptococcus, Collinsella, Prevotella, Parvimonas, and Gemella, in patients with CRC. An integrated analysis using 16S rRNA data and epidemiological characteristics showed that alcohol consumption was significantly correlated with the abundance of Fusobacterium OTUs. The correlation of alcohol consumption with the abundance of Fusobacterium OTUs in cancer tissue discovered using 16S rRNA analysis suggests a possible link between alcohol metabolism and subsequent tumorigenesis caused by F. nucleatum.

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Section: Methodsmentioning

confidence: 79%

“…1C). Thus, it may be necessary to evaluate both F. nucleatum presence and fadA gene frequency to accurately determine the presence and virulence of F. nucleatum 31 . Interestingly, a recent study suggested that the fadA gene is not specific to F. nucleatum and that it is also present in other Fusobacterium species 32 .…”

Section: Discussionmentioning

confidence: 99%

Fusobacterium nucleatum in biopsied tissues from colorectal cancer patients and alcohol consumption in Korea

Kim

Lee

Choi

et al. 2020

Sci Rep

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“…Fecal-based F. nucleatum polymerase chain reaction (PCR) can serve as a noninvasive tool for CRC detection, with even better results when using digital PCR based on water-oil emulsion droplet technology[ 39 , 54 , 106 - 109 ]. Compared with PCR, loop-mediated isothermal amplification (LAMP) is a simple, noncostly and accurate method for bacterial testing that was shown to be more sensitive than PCR for F. nucleatum detection[ 110 ]. Two drawbacks of LAMP are the potential for false positivity and the complex design primer used.…”

Section: Resultsmentioning

confidence: 99%

Fusobacterium’s link to colorectal neoplasia sequenced: A systematic review and future insights

Hussan¹,

Clinton²,

Roberts³

et al. 2017

WJG

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AIMTo critically evaluate previous scientific evidence on Fusobacterium’s role in colorectal neoplasia development.METHODSTwo independent investigators systematically reviewed all original scientific articles published between January, 2000, and July, 2017, using PubMed, EMBASE, and MEDLINE. A total of 355 articles were screened at the abstract level. Of these, only original scientific human, animal, and in vitro studies investigating Fusobacterium and its relationship with colorectal cancer (CRC) were included in the analysis. Abstracts, review articles, studies investigating other colonic diseases, and studies written in other languages than English were excluded from our analysis. Ninety articles were included after removing duplicates, resolving disagreements between the two reviewers, and applying the above criteria.RESULTSStudies have consistently identified positive associations between Fusobacterium, especially Fusobacterium nucleatum (F. nucleatum), and CRC. Stronger associations were seen in CRCs proximal to the splenic flexure and CpG island methylator phenotype (CIMP)-high CRCs. There was evidence of temporality and a biological gradient, with increased F. nucleatum DNA detection and quantity along the traditional adenoma-carcinoma sequence and in CIMP-high CRC precursors. Diet may have a differential impact on colonic F. nucleatum enrichment; evidence suggests that high fiber diet may reduce the risk of a subset of CRCs that are F. nucleatum DNA-positive. Data also suggest shorter CRC and disease-specific survival with increased amount of F. nucleatum DNA in CRC tissue. The pathophysiology of enrichment of F. nucleatum and other Fusobacterium species in colonic tissue is unclear; however, the virulence factors and changes to the local colonic environment with disruption of the protective mucus layer may contribute. The presence of a host lectin (Gal-GalNAc) in the colonic epithelium may also mediate F. nucleatum attachment to CRC and precursors through interaction with an F. nucleatum protein, fibroblast activation protein 2 (FAP2). The clinical significance of detection or enrichment of Fusobacterium in colorectal neoplasia is ambiguous, but data suggest a procarcinogenic effect of F. nucleatum, likely due to activation of oncogenic and inflammatory pathways and modulation of the tumor immune environment. This is hypothesized to be mediated by certain F. nucleatum strains carrying invasive properties and virulence factors such as FadA and FAP.CONCLUSIONEvidence suggests a potential active role of Fusobacterium, specifically F. nucleatum, in CRC. Future prospective and experimental human studies would fill an important gap in this literature.

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“…The detection target is highly conserved nusG gene of Fusobacterium nucleatum [17]. The size of the rst-and second-stage PCR products is expected at 175 and 124 bp, respectively.…”

Section: Resultsmentioning

confidence: 99%

Fusobacterium nucleatum Promotes Gastric Cancer Aggressiveness through Upregulation of Cell Mobility and Interferon Genes

Hsieh

Tung

Pan

et al. 2020

Preprint

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Background Fusobacterium nucleatum was previously found to become a dominant species in the gastric cancer-associated microbiota of patients from Taiwan. However, the prevalence of Fusobacterium nucleatum infection in gastric cancer has not been examined in a larger patient cohort. In addition, whether Fusobacterium nucleatum elicits a cellular response in gastric cancer remains unknown.Methods A study cohort of resected gastric cancer tissue specimens was examined using nested PCR to detect Fusobacterium nucleatum. In vitro coculture of Fusobacterium nucleatum was carried out to identify the alteration in the expression profile of patient-derived gastric cancer cell line.Results approximately one-third of gastric cancer tissues are positive for Fusobacterium nucleatum. Statistical analysis showed that the risk for Fusobacterium nucleatum infection is increased in late-stage cancer tissue specimens and incurs poorer survival in Helicobacter pylori-positive patients. In vitro coculture experiment shows a drastic interferon response activated only by a high multiplicity of infection, and the response peaks within 24 hours and subsides after 72 hours of incubation. Another set of response genes is the continuous increase of actins and their regulators with prolonged time of incubation, activated by both low and high multiplicity of infection.Conclusions Our data indicates that Fusobacterium nucleatum incites an inflammatory response from the cancer cells and promotes cell mobility, likely

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Rapid detection of nusG and fadA in Fusobacterium nucleatum by loop-mediated isothermal amplification

Cited by 15 publications

References 27 publications

Fusobacterium nucleatum in biopsied tissues from colorectal cancer patients and alcohol consumption in Korea

Fusobacterium nucleatum in biopsied tissues from colorectal cancer patients and alcohol consumption in Korea

Fusobacterium’s link to colorectal neoplasia sequenced: A systematic review and future insights

Fusobacterium nucleatum Promotes Gastric Cancer Aggressiveness through Upregulation of Cell Mobility and Interferon Genes

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