[Establishment of loop-mediated isothermal amplification for detection of Vibrio alginolyticus].
Vibrio alginolyticus is the main Vibrio pathogen in aquaculture in the south of China. A one step loop-mediated isothermal amplification (LAMP) assay was developed for detection of Vibrio alginolyticus. A set of primers were designed from the OmpK sequence of Vibrio alginolyticus. The assay was optimised to amplify Vibrio alginolyticus DNA by incubation at 65 degrees C for only 1 h, and required only a simple water bath or heating block to provide a constant temperature of 65 degrees C. LAMP amplification products had a ladder-like appearance when electrophoresed on an agarose gel. The detection limit of the LAMP assay was n(cell) = 38/mL which was found to be higher than the commonly used PCR method. The assay was evaluated using clinical samples and the results indicated the suitability and simplicity of the test as a rapid, field diagnostic tool for Vibrio alginolyticus.
Ding
Wen Chao
WC
Key Laboratory of Applied Marine Biotechnology, Ministry of Education, Ningbo University, Ningbo 315211.
Hu
Jian Rao
JR
Shi
Yu Hong
YH
Li
Ming Yun
MY
Chen
Jiong
J
D001431:Bacteriological Techniques / Q000379:methods*; D004269:DNA, Bacterial / Q000235:genetics; D021141:Nucleic Acid Amplification Techniques / Q000379:methods*; D016133:Polymerase Chain Reaction; D015203:Reproducibility of Results; D013696:Temperature; D044149:Vibrio alginolyticus / Q000235:genetics* / Q000302:isolation & purification
D004269:DNA, Bacterial