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Evaluation of a loop-mediated isothermal amplification method for the rapid detection of Vibrio harveyi in cultured marine shellfish
Cao, YT; Wu, ZH; Jian, JC; Lu, YS; wuzh@gdou.edu.cn
2010
发表期刊LETTERS IN APPLIED MICROBIOLOGY
ISSN0266-8254
卷号51期号:1页码:24-29
摘要Aims: The purpose of this study was to establish a loop-mediated isothermal amplification (LAMP) method for the rapid, sensitive detection of Vibrio harveyi in mariculture shellfish. Methods and Results: A set of four primers, two outer and two inner primers, were designed from the toxR gene sequence of V. harveyi. The LAMP reaction was conducted at 65 degrees C for 60 min. There were no cross-reactions with other bacterial strains indicating a high specificity of the LAMP. The detection sensitivity of the LAMP assay for V. harveyi with both of pure cultures and added shellfish cultures is about 10-5 dilution level (equivalent to 17 center dot 2 cells per reaction). The amplification products were detected by visual inspection using SYBR Green I. The detection sensitivity using the LAMP method was 10 times higher than that of conventional PCR. Conclusions: The LAMP assay established in this study is an extremely specific, sensitive and rapid for identification of V. harveyi in mariculture shellfish. Significance and Impact of the Study: This LAMP technique provides an important detecting tool for the detection of V. harveyi infection both in the laboratory and field. This technique is recommended as an applied protocol for health management programme and disease surveillance of in hatcheries as well as in grow-out pond, to prevent the disease outbreak.
部门归属[Cao, Y. T.; Wu, Z. H.; Jian, J. C.; Lu, Y. S.] Guangdong Ocean Univ, Coll Fisheries, Zhanjiang, Peoples R China; [Cao, Y. T.; Wu, Z. H.; Jian, J. C.; Lu, Y. S.] Chinese Acad Sci, S China Sea Inst Oceanol, Guangzhou, Guangdong, Peoples R China; [Cao, Y. T.; Wu, Z. H.; Jian, J. C.; Lu, Y. S.] Guangdong Prov Key Lab Pathogen Biol & Epidemiol, Zhanjiang, Peoples R China; [Cao, Y. T.] Chinese Acad Sci, Grad Sch, Beijing, Peoples R China
关键词Detection Loop-mediated Isothermal Amplification Toxr Gene Vibrio Harveyi
学科领域Biotechnology & Applied Microbiology ; Microbiology
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收录类别sci
资助项目海洋生物
WOS记录号WOS:000278623300005
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被引频次:11[WOS]   [WOS记录]     [WOS相关记录]
文献类型期刊论文
条目标识符http://ir.scsio.ac.cn/handle/344004/5426
专题海洋生物
通讯作者wuzh@gdou.edu.cn
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Cao, YT,Wu, ZH,Jian, JC,et al. Evaluation of a loop-mediated isothermal amplification method for the rapid detection of Vibrio harveyi in cultured marine shellfish[J]. LETTERS IN APPLIED MICROBIOLOGY,2010,51(1):24-29.
APA Cao, YT,Wu, ZH,Jian, JC,Lu, YS,&wuzh@gdou.edu.cn.(2010).Evaluation of a loop-mediated isothermal amplification method for the rapid detection of Vibrio harveyi in cultured marine shellfish.LETTERS IN APPLIED MICROBIOLOGY,51(1),24-29.
MLA Cao, YT,et al."Evaluation of a loop-mediated isothermal amplification method for the rapid detection of Vibrio harveyi in cultured marine shellfish".LETTERS IN APPLIED MICROBIOLOGY 51.1(2010):24-29.
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