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

Abstract:

AIMS: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. METHODS AND RESULTS: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 degrees C in a simple water bath. The detection sensitivity of the LAMP assay for the detection of V. alginolyticus is about 3.7 x 10(2) 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. CONCLUSIONS:The LAMP established in this study is a simple, sensitive, specific, inexpensive and rapid protocol for the detection of V. alginolyticus. SIGNIFICANCE AND IMPACT OF THE STUDY:This LAMP method provides an important diagnostic tool for the detection of V. alginolyticus infection both in the laboratory and field.

journal_name

Lett Appl Microbiol

authors

Cai SH,Lu YS,Wu ZH,Jian JC,Wang B,Huang YC

doi

10.1111/j.1472-765X.2010.02823.x

subject

Has Abstract

pub_date

2010-05-01 00:00:00

pages

480-5

issue

5

eissn

0266-8254

issn

1472-765X

pii

LAM2823

journal_volume

50

pub_type

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