title: Rapid detection of nusG and fadA in Fusobacterium nucleatum by loop-mediated isothermal amplification.
abstract: Fusobacterium nucleatum is associated with various human diseases such as periodontal disease and colorectal cancer (CRC); thus, F. nucleatum detection might serve as a novel diagnostic tool. Here, we describe the development of a sensitive and rapid molecular method for detecting two F. nucleatum genes: the highly conserved nusG and fadA, which encode a critical host colonization factor. Loop-mediated isothermal amplification (LAMP) primer sets for the rapid detection of nusG and fadA were designed and optimized. The nusG primers yielded consistent negative results for 20 non-F. nucleatum bacterial strains, confirming the high specificity of the primers. LAMP reaction primer sensitivity was determined, and its detection rate in comparison to conventional PCR was assessed using 57 clinical stool samples. The LAMP detection limit for nusG and fadA was 22.5 and 0.225 pg µl-1, respectively, indicating that the sensitivity of this method was 10-fold higher than that of conventional PCR. These results suggest that the LAMP technique is able to effectively identify F. nucleatum via nusG as well as detect its virulence factor. To the best of our knowledge, this study is the first to report the application of LAMP for the detection of nusG and fadA in F. nucleatum. The LAMP method constitutes a sensitive and specific visual assay for the rapid detection of the pathogen F. nucleatum.
authors.affiliation: Institute of Disease Control and Prevention, Academy of Military Medical Sciences, Beijing, P. R. China.
authors.forename: Simo
authors.initials: S
authors.lastname: Huang
authors.affiliation: Institute of Disease Control and Prevention, Academy of Military Medical Sciences, Beijing, P. R. China.
authors.forename: Zhan
authors.initials: Z
authors.lastname: Yang
authors.affiliation: Institute of Disease Control and Prevention, Academy of Military Medical Sciences, Beijing, P. R. China.
authors.forename: Dayang
authors.initials: D
authors.lastname: Zou
authors.affiliation: Institute of Disease Control and Prevention, Academy of Military Medical Sciences, Beijing, P. R. China.
authors.forename: Derong
authors.initials: D
authors.lastname: Dong
authors.affiliation: Institute of Disease Control and Prevention, Academy of Military Medical Sciences, Beijing, P. R. China.
authors.forename: Anheng
authors.initials: A
authors.lastname: Liu
authors.affiliation: Institute of Disease Control and Prevention, Academy of Military Medical Sciences, Beijing, P. R. China.
authors.forename: Wei
authors.initials: W
authors.lastname: Liu
authors.affiliation: Institute of Disease Control and Prevention, Academy of Military Medical Sciences, Beijing, P. R. China.
authors.forename: Liuyu
authors.initials: L
authors.lastname: Huang
mesh_terms: D000328:Adult; D000368:Aged; D001426:Bacterial Proteins / Q000235:genetics*; D017931:DNA Primers / Q000235:genetics; D005260:Female; D005674:Fusobacterium Infections / Q000175:diagnosis*; D016967:Fusobacterium nucleatum / Q000235:genetics / Q000302:isolation & purification*; D006801:Humans; D008297:Male; D008875:Middle Aged; D025202:Molecular Diagnostic Techniques / Q000379:methods*; D021141:Nucleic Acid Amplification Techniques / Q000379:methods*; D012680:Sensitivity and Specificity; D013997:Time Factors
chemical_list: D001426:Bacterial Proteins; D017931:DNA Primers