title: A rapid, simple, and sensitive loop-mediated isothermal amplification method to detect toxigenic Vibrio cholerae in rectal swab samples.
abstract: Loop-mediated isothermal amplification (LAMP) method was designed for clinical diagnosis of Vibrio cholerae carrying the ctxA gene. The detection limits of the method were 5 fg of purified genomic DNA/reaction and 0.54 CFU/reaction. The method was applied to rectal swab samples from cholera patients and healthy volunteers (19 subjects each) and yielded the same results as the "gold standard" culture method, while the polymerase chain reaction-based method failed to detect V. cholerae in 8 of the positive samples. Direct application of this LAMP method without precultivation enabled the rapid detection of 5 asymptomatic carriers from rectal swabs of 21 household contacts of cholera patients. This LAMP method could be a sensitive, specific, inexpensive, and rapid detection tool for V. cholerae carrying the ctxA gene in the clinical laboratory and in the field.
authors.affiliation: Thailand-Japan Research Collaboration Center on Emerging and Reemerging Infections, DMSc, Ministry of Public Health, Nonthaburi, Thailand. kazuhisa@biken.osaka-u.ac.jp
authors.forename: Kazuhisa
authors.initials: K
authors.lastname: Okada
authors.forename: Siriporn
authors.initials: S
authors.lastname: Chantaroj
authors.forename: Tooru
authors.initials: T
authors.lastname: Taniguchi
authors.forename: Yasuhiko
authors.initials: Y
authors.lastname: Suzuki
authors.forename: Amonrattana
authors.initials: A
authors.lastname: Roobthaisong
authors.forename: Orapim
authors.initials: O
authors.lastname: Puiprom
authors.forename: Takeshi
authors.initials: T
authors.lastname: Honda
authors.forename: Pathom
authors.initials: P
authors.lastname: Sawanpanyalert
mesh_terms: D001426:Bacterial Proteins / Q000235:genetics*; D002353:Carrier State / Q000175:diagnosis / Q000382:microbiology; D002771:Cholera / Q000175:diagnosis* / Q000382:microbiology; D002772:Cholera Toxin / Q000235:genetics*; D019411:Clinical Laboratory Techniques / Q000379:methods*; D006801:Humans; D021141:Nucleic Acid Amplification Techniques / Q000379:methods*; D012007:Rectum / Q000382:microbiology*; D012680:Sensitivity and Specificity; D014734:Vibrio cholerae / Q000235:genetics / Q000302:isolation & purification*
chemical_list: D001426:Bacterial Proteins; D002772:Cholera Toxin