Rapid detection of FadA in Fusobacterium nucleatum using the quantitative LAMP colorimetric phenol red method in stool samples from colorectal cancer patients.
Humans
Colorectal Neoplasms
/ microbiology
Fusobacterium nucleatum
/ genetics
Feces
/ microbiology
Nucleic Acid Amplification Techniques
/ methods
Colorimetry
/ methods
Male
Female
Phenolsulfonphthalein
Molecular Diagnostic Techniques
/ methods
Middle Aged
Aged
Fusobacterium Infections
/ microbiology
Sensitivity and Specificity
Adult
FadA
Fusobacterium nucleatum
Colorectal cancer (CRC)
Phenol red
QLAMP
Journal
Scientific reports
ISSN: 2045-2322
Titre abrégé: Sci Rep
Pays: England
ID NLM: 101563288
Informations de publication
Date de publication:
14 Jun 2024
14 Jun 2024
Historique:
received:
07
11
2023
accepted:
22
05
2024
medline:
15
6
2024
pubmed:
15
6
2024
entrez:
14
6
2024
Statut:
epublish
Résumé
The study aimed to develop a quantitative colorimetric loop-mediated isothermal amplification technique using the phenol red indicator (QLAMP-PhR) for detecting Fusobacterium nucleatum (Fn) levels in colorectal cancer (CRC) patients and healthy individuals. QLAMP-PhR assays were conducted on 251 stool samples specific for the Fn FadA gene. Six primers were synthesized and utilized with master mix reagents, and a phenol red indicator was employed to enhance the QLAMP-PhR technique. A standard quantitative analysis curve was generated using a logarithmic function (absorbance vs. concentration) by serially diluting the copy number of genomic DNA templates (Fn ATCC25586). The CRC group exhibited a significantly higher abundance of Fn compared to the healthy control group (P < 0.001). These findings suggest that the QLAMP-PhR technique effectively identifies Fn specifically by its gene for the key virulence factor FadA. Additionally, ideas for developing a real-time QLAMP-PhR test were presented. Compared to the traditional polymerase chain reaction (PCR) technique, QLAMP-PhR offers several advantages including rapidity, simplicity, specificity, sensitivity, and cost-effectiveness method that can quantitatively screen for Fn presence in normal populations. The QLAMP-PhR method represents a sensitive and specific amplification assay for the rapid detection of the Fn pathogen. To the best of our knowledge, this study is the first to report the application of QLAMP-PhR for detecting FadA in Fn.
Identifiants
pubmed: 38877111
doi: 10.1038/s41598-024-62846-x
pii: 10.1038/s41598-024-62846-x
doi:
Substances chimiques
Phenolsulfonphthalein
I6G9Y0J1OJ
Types de publication
Journal Article
Langues
eng
Sous-ensembles de citation
IM
Pagination
13739Informations de copyright
© 2024. The Author(s).
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