A rapid, simple, and sensitive loop-mediated isothermal amplification method to detect toxigenic Vibrio cholerae in rectal swab samples.
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.
Okada
Kazuhisa
K
Thailand-Japan Research Collaboration Center on Emerging and Reemerging Infections, DMSc, Ministry of Public Health, Nonthaburi, Thailand. kazuhisa@biken.osaka-u.ac.jp
Chantaroj
Siriporn
S
Taniguchi
Tooru
T
Suzuki
Yasuhiko
Y
Roobthaisong
Amonrattana
A
Puiprom
Orapim
O
Honda
Takeshi
T
Sawanpanyalert
Pathom
P
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*
D001426:Bacterial Proteins; D002772:Cholera Toxin