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Loop-mediated isothermal amplification for rapid detection of Acute viral necrobiotic virus in scallop Chlamys farreii

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文献类型：外文期刊

作者： Ren, W. C. ¹ ; Wang, C. M. ¹ ; Cai, Y. Y. ¹ ;

作者机构： 1.Chinese Acad Fishery Sci, Maricultural Organism Dis Control & Pathogen Mol, Yellow Sea Fisheries Res Inst, Qingdao 266071, Shandong, Peoples R China

2.Ocean Univ China, Dept Aquaculture, Coll Fisheries, Qingdao 266003, Shandong, Peoples R China

关键词： Acute viral necrobiotic virus;scallop Chlamys farreri;loop-mediated isothermal amplification

期刊名称：ACTA VIROLOGICA （影响因子：1.162；五年影响因子：1.255 ）

ISSN： 0001-723X

年卷期： 2009 年 53 卷 3 期

页码：

收录情况： SCI

摘要： Loop-mediated isothermal amplification (LAMP) assay for rapid and sensitive detection of Acute viral necrobiotic virus (AVNV) in scallop Chlamys farreri was developed and evaluated. Four primers recognizing six targets on distinct AVNV DNA sequences were designed and the LAMP reaction was carried out in a water bath. Reaction temperature and time were optimized at 64 degrees C for 60 mins and LAMP products were detected using agarose gel electrophoresis and visual assessment. Confirmation of the expected LAMP products was performed with MboI restriction enzyme analysis. The detection limit of LAMP assay was as low as 1 fg AVNV DNA and accordingly, this assay was 100 times more sensitive than conventional PCR technique. A comparative evaluation of 20 samples using the LAMP and PCR assays revealed a complete accord in positivity or negativity for AVNV. These results indicate that the LAMP assay is simple, sensitive, specific, and has a great potential for detection of AVNV in the laboratory and field.

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Loop-mediated isothermal amplification for rapid detection of Acute viral necrobiotic virus in scallop Chlamys farreii

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Loop-mediated isothermal amplification for rapid detection of Acute viral necrobiotic virus in scallop Chlamys farreii

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