Sensitive and rapid detection of Vibrio corallilyticus by loop-mediated isothermal amplification targeted to the alpha subunit gene of RNA polymerase
文献类型: 外文期刊
作者: Liu, G. F. 1 ; Wang, J. Y. 2 ; Xu, L. W. 2 ; Ding, X. 2 ; Zhou, S. N. 1 ;
作者机构: 1.Sun Yat Sen Univ, Sch Life Sci, Guangzhou 510275, Guangdong, Peoples R China
2.Chinese Acad Fishery Sci, S China Sea Fisheries Res Inst, Guangzhou, Guangdong, Peoples R China
关键词: detection.;loop-mediated isothermal amplification;PCR;rpo A gene;Vibrio corallilyticus;Internet resource.
期刊名称:LETTERS IN APPLIED MICROBIOLOGY ( 影响因子:2.858; 五年影响因子:2.776 )
ISSN:
年卷期:
页码:
收录情况: SCI
摘要: A diagnostic protocol was developed for rapid detection of Vibrio corallilyticus by method of loop-mediated isothermal amplification (LAMP). For cloning and sequencing of rpo A gene of V. corallilyticus, a set of four LAMP primers were designed by targeting the rpoA gene. With Bst DNA polymerase, the reaction time and temperature were optimized for 70 min at 65pC, respectively. The amplification products were detected by electrophoresis. The detection limit of V. corallilyticus by LAMP was 3p"6x10pd CFU mlp# (8 CFU per reaction), but PCR could detect up to 3p"6x10t CFU mlp# (72 CFU per reaction). The LAMP method was ninefold more sensitive than conventional PCR. The results also indicated that the LAMP reaction was highly specific to V. corallilyticus. The LAMP assay was a sensitive, specific and cost-effective method for the rapid detection of V. corallilyticus. This LAMP method provides an important diagnostic tool for the detection of V. corallilyticus infection. It can replace laborious biochemical tests for the identification of V. corallilyticus.
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