[Establishment and application of a loop-mediated isothermal amplification method for rapid diagnosis of Vibrio cholerae].
OBJECTIVE
To establish a loop-mediated isothermal amplification (LAMP) method for rapid diagnosis of Vibrio cholerae.


METHODS
Based on the ompW nucleic sequence of Vibrio cholerae, a pair of primers was designed for LAMP. The reaction conditions were optimized, and the specificity, sensitivity, and practicability of LAMP were tested using 47 bacterial strains and simulated contaminated sites.


RESULTS
The results of viable bacterium count showed that LAMP was capable of detecting Vibrio cholerae at a level as low as 1.6x10(2) cfu/ml. The minimal detectable concentration was 1.6+10(3) cfu/ml for simulated contaminated samples such as feces and seawater, and 1.6+10(4) cfu/ml for contaminated milk. All the 21 strains of Vibrio cholerae yielded positive results in LAMP, and the 26 strains of other bacteria all showed negative results, with a detection specificity of 100%.


CONCLUSIONS
The established LAMP method has high specificity and sensitivity for detecting Vibrio cholerae and is applicable in field monitoring and epidemiological study of Vibrio cholerae.
Ke
Xue-mei
XM
School of Public Health and Tropical Medicine, Southern Medical University, Guangzhou 510515, China. xmke023@126.com
Chen
Yin-yu
YY
Gao
Lu-lu
LL
DU
Zheng-ping
ZP
Feng
Xue-mei
XM
Liao
Ru-yan
RY
Chen
Zhi-yong
ZY
Cao
Yi-cheng
YC
Chen
Qing
Q
D001426:Bacterial Proteins / Q000235:genetics; D002771:Cholera / Q000175:diagnosis* / Q000382:microbiology; D019411:Clinical Laboratory Techniques / Q000379:methods*; D006801:Humans; D021141:Nucleic Acid Amplification Techniques / Q000379:methods*; D012680:Sensitivity and Specificity; D014734:Vibrio cholerae / Q000235:genetics / Q000302:isolation & purification*
D001426:Bacterial Proteins