สถาบันวิจัยวิทยาศาสตร์สาธารณสุข

NATIONAL INSTITUTE OF HEALTH OF THAILAND

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

Authors : Kazuhisa Okadaa, Siriporn Chantarojb, Tooru Taniguchia, Yasuhiko Suzukic, Amonrattana Roobthaisonga, Orapim Puiproma, Takeshi Hondaa, Pathom Sawanpanyalertb

 

Affiliations : 

a Section of Bacterial Infections, Thailand–Japan Research Collaboration Center on Emerging and Reemerging Infections (RCC-ERI), DMSc, Ministry of Public Health, Nonthaburi 11000, Thailand
b National Institute of Health, DMSc, Tiwanond Road, Nonthaburi 11000, Thailand
c Department of Global Epidemiology, Hokkaido University Research Center for Zoonosis Control, N20, W10, Kita-ku, Sapporo, Hokkaido 001-0020, Japan. (Received 17 April 2009; accepted 2 September 2009)

Source :

Elsevier Inc. Diagnostic Microbiology and Infectious Disease 2010; 135-139  

Language : English

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.