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

被引：41

作者：

Cai, S. H. ^{[1
,2
]}

Lu, Y. S. ^{[1
,2
]}

Wu, Z. -H. ^{[1
,2
]}

Jian, J. C. ^{[1
,2
]}

Wang, B. ^{[1
,2
]}

Huang, Y. C. ^{[1
,2
]}

机构：

[1] Guangdong Ocean Univ, Coll Fisheries, Zhanjiang 524025, Guangdong, Peoples R China

[2] Guangdong Prov Key Lab Pathogen Biol & Epidemiol, Zhanjiang, Peoples R China

来源：

LETTERS IN APPLIED MICROBIOLOGY | 2010年 / 50卷 / 05期

关键词：

loop-mediated isothermal amplification; PCR; rapid detection; Vibrio alginolyticus; GYRB GENE; PCR; PARAHAEMOLYTICUS; SHRIMP; SEQUENCE; SEAWATER; CLONING;

D O I：

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

中图分类号：

Q81 [生物工程学（生物技术）]; Q93 [微生物学];

学科分类号：

071005 ; 0836 ; 090102 ; 100705 ;

摘要：

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 center dot 7 x 102 CFU ml-1 (3 center dot 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.

引用

页码：480 / 485

页数：6

共 21 条

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