Rapid detection of a fish iridovirus using loop-mediated isothermal amplification (LAMP)
Rapid detection of a fish iridovirus using loop-mediated isothermal amplification (LAMP)
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DOI：
10.1016/j.jviromet.2004.06.011
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发表时间：
2004-11-01
影响因子：
3.1
通讯作者：
Aoki, T
中科院分区：
文献类型：
--
作者：
Caipang, CMA;Haraguchi, I;Aoki, T
采用环介导的等温扩增（LAMP）方法检测红海鲷虹膜病毒（RSIV）的基因组DNA分子。RSIV是虹膜病毒科的一种鱼类虹膜病毒。从感染的红鲷、大黄鲷和各种RSIV分离株的脾脏中提取DNA，扩增RSIV DNA。该方法检测RSIV的灵敏度是传统PCR的10倍以上。LAMP反应的一个显著特征是它能够合成大量的DNA，从而产生一种白色沉淀物——焦磷酸镁——作为副产品。这种白色沉淀物的存在或不存在有助于不使用凝胶电泳的RSIV基因组DNA的检测。在反应结束时，输入的病毒DNA拷贝量与相应的浊度读数之间存在很强的相关性；因此，LAMP反应可能用于定量感染鱼体内的RSIV颗粒。(C) 2004 Elsevier B.V.版权所有
A loop-mediated isothermal amplification (LAMP) procedure is described to detect the genomic DNA molecule of red seabream iridovirus (RSIV), a fish iridovirus belonging to the Iridoviridae family. The RSIV DNA was amplified using DNA extracts obtained from spleen of infected red seabream, Pagrus major and from various RSIV isolates. The method was at least 10 times more sensitive than conventional PCR in detecting for the presence of RSIV. A striking feature of the LAMP reaction is its ability to synthesize a large amount of DNA leading to the production of a white precipitate, magnesium pyrophosphate, as a by-product. The presence or absence of this white precipitate facilitates easy detection of the RSIV genomic DNA without the use of gel electrophoresis. A strong correlation exists between the amount of input viral DNA copy and the corresponding turbidity reading at the end of the reaction; hence, the LAMP reaction may be used potentially to quantify RSIV particles in the infected fish. (C) 2004 Elsevier B.V. All rights reserved.