Establishment and application of a loop-mediated isothermal amplification method for rapid diagnosis of Vibrio cholerae.
- Author:
Xue-mei KE
1
;
Yin-yu CHEN
;
Lu-lu GAO
;
Zheng-ping DU
;
Xue-mei FENG
;
Ru-yan LIAO
;
Zhi-yong CHEN
;
Yi-cheng CAO
;
Qing CHEN
Author Information
- Publication Type:Journal Article
- MeSH: Bacterial Proteins; genetics; Cholera; diagnosis; microbiology; Clinical Laboratory Techniques; methods; Humans; Nucleic Acid Amplification Techniques; methods; Sensitivity and Specificity; Vibrio cholerae; genetics; isolation & purification
- From: Journal of Southern Medical University 2009;29(10):2059-2063
- CountryChina
- Language:Chinese
-
Abstract:
OBJECTIVETo establish a loop-mediated isothermal amplification (LAMP) method for rapid diagnosis of Vibrio cholerae.
METHODSBased 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.
RESULTSThe 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%.
CONCLUSIONThe established LAMP method has high specificity and sensitivity for detecting Vibrio cholerae and is applicable in field monitoring and epidemiological study of Vibrio cholerae.
