Development of a loop-mediated isothermal amplification assay for detecting Vibrio parahaemolyticus.
- Author:
Xin LU
1
;
Shu-jing WANG
;
Sha LIU
;
Biao KAN
;
Bo PANG
Author Information
- Publication Type:Journal Article
- MeSH: Nucleic Acid Amplification Techniques; methods; Sensitivity and Specificity; Vibrio parahaemolyticus; genetics; isolation & purification
- From: Chinese Journal of Preventive Medicine 2012;46(5):465-467
- CountryChina
- Language:Chinese
-
Abstract:
OBJECTIVEThis study aimed to establish a loop-mediated isothermal amplification (LAMP) assay for detection of Vibrio parahaemolyticus (V. parahaemolyticus).
METHODSThe specificity of this assay was evaluated by using a panel of 33 strains of V. parahaemolyticus and 22 strains of other species bacteria. The sensitivity was determined by using serial dilutions of V. parahaemolyticus (ATCC 17802) chromosomal DNA (5×10(0) - 5×10(5) copies/µl). The samples were also tested by using qualification PCR assay and Taqman real-time PCR assay in parallel for comparison with LAMP.
RESULTSBoth sensitivity and specificity of LAMP assay, PCR assay and Taqman real-time PCR assay were 100% (22/22, 33/33, respectively). The detection limits of above three methods assay were 5×10(1) copies/µl, 5×10(3) copies/µl and 5×10(2) copies/µl, respectively. The reaction period of time needed of the above three assays was 22 min, 3 h, 50 min, respectively.
CONCLUSIONCompared to qualification PCR assay and Taqman real-time PCR assay, the established LAMP assay was better in low detection limit and less reaction time, which made it an ideal method for quick detection of V. parahaemolyticus.