A rapid, simple, and sensitive loop-mediated isothermal amplification method to detect toxigenic Vibrio cholerae in rectal swab samples

作者：Okada Kazuhisa^*; Chantaroj Siri****; Taniguchi Tooru; Suzuki Yasuhiko; Roobthaisong Amonrattana; Puiprom Orapim; Honda Takeshi; Sawanpanyalert Pathom

来源：Diagnostic Microbiology and Infectious Disease, 2010, 66(2): 135-139.

DOI：10.1016/j.diagmicrobio.2009.09.004

• Vibrio cholerae
• LAMP
• ctxA
• PCR
• Cholera

Loop-mediated isothermal amplification (LAMP) method was designed for clinical diagnosis of Vibrio cholerae carrying the call 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.

• 出版日期2010-2