Loop-mediated isothermal amplification method for rapid detection of Vibrio alginolyticus, the causative agent of vibriosis in mariculture fish

The purpose of this study was to develop a loop-mediated isothermal amplification (LAMP) method for the rapid, sensitive and simple detection of Vibrio alginolyticus in mariculture fish. LAMP primers were designed by targeting the gyrB gene. With Bst DNA polymerase, the target DNA can be clearly amp...

Full description

Saved in:
Bibliographic Details
Published in:Letters in applied microbiology 2010-05, Vol.50 (5), p.480-485
Main Authors: Cai, S.H, Lu, Y.S, Wu, Z.-H, Jian, J.C, Wang, B, Huang, Y.C
Format: Article
Language:English
Subjects:
Animals
Bacterial Proteins - genetics
DNA Primers - genetics
Fish Diseases - microbiology
loop-mediated isothermal amplification
Nucleic Acid Amplification Techniques - methods
PCR
rapid detection
Vibrio alginolyticus
Vibrio alginolyticus - genetics
Vibrio alginolyticus - isolation & purification
Vibrio Infections - microbiology
Vibrio Infections - veterinary
Citations: Items that this one cites
Items that cite this one
Online Access:Get full text
Tags: Add Tag
No Tags, Be the first to tag this record!
Description
Summary:The purpose of this study was to develop a loop-mediated isothermal amplification (LAMP) method for the rapid, sensitive and simple detection of Vibrio alginolyticus in mariculture fish. LAMP primers were designed by targeting the gyrB gene. With Bst DNA polymerase, the target DNA can be clearly amplified for 60 min at 64°C in a simple water bath. The detection sensitivity of the LAMP assay for the detection of V. alginolyticus is about 3·7 x 10² CFU ml⁻¹ (3·7 CFU per reaction). LAMP products could be judged with agar gel or naked eye after the addition of SYBR Green I. There were no cross-reactions with other bacterial strains indicating a high specificity of the LAMP. The LAMP method was applied to detect V. alginolyticus-infected fish tissues effectively. The LAMP established in this study is a simple, sensitive, specific, inexpensive and rapid protocol for the detection of V. alginolyticus. This LAMP method provides an important diagnostic tool for the detection of V. alginolyticus infection both in the laboratory and field.
ISSN:0266-8254
1472-765X
DOI:10.1111/j.1472-765X.2010.02823.x