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 Microbioljournal_title
Letters in applied microbiologyauthors
Cai SH,Lu YS,Wu ZH,Jian JC,Wang B,Huang YCdoi
10.1111/j.1472-765X.2010.02823.xsubject
Has Abstractpub_date
2010-05-01 00:00:00pages
480-5issue
5eissn
0266-8254issn
1472-765Xpii
LAM2823journal_volume
50pub_type
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