Easy-to-use rapid gene amplification method for direct detection of RNA and DNA viruses in sera and feces from various animals
Easy-to-use rapid gene amplification method for direct detection of RNA and DNA viruses in sera and feces from various animals
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DOI：
10.1016/j.jviromet.2014.01.019
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发表时间：
2014-06-01
影响因子：
3.1
通讯作者：
Sakai, Takeo
中科院分区：
文献类型：
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作者：
Segawa, Takao;Kobayashi, Yuki;Sakai, Takeo
需要开发快速和简单的基因扩增测试来检测病原体，以防止动物之间或从动物到人类的传染病传播。建立了一种简便快速的基因扩增方法，可直接检测临床标本中的RNA和DNA病毒。该方法基于环介导等温扩增（LAMP）或逆转录-LAMP（RT-LAMP）和RNA GEM Tissue（一种通过快速消化蛋白质和核糖核酸酶提取核酸的嗜热酶）的组合。作者将这些方法命名为GEM LAMP和GEM RT-LAMP。这些方法能够在70分钟内检测病毒的DNA和RNA在一个单一的管中使用只有一个水浴。检测能力比先前建立的LAMP和RT-LAMP方法灵敏10-100倍。GEM LAMP和GEM RT-LAMP方法用于使用SYBR绿色I肉眼检测牛、猪、马、海豚、企鹅和海狮血清或粪便样本中是否存在DNA和RNA病毒。因此，GEM LAMP和GEM RT-LAMP方法作为用于检测病原体的工具具有相当大的通用性，并且适用于基本的人类和兽医学、环境卫生和护理点测试。(C)2014爱思唯尔有限公司版权所有。
The development of rapid and simple gene amplification tests is required for detection of pathogens to prevent transmission of infectious diseases between animals or from animals to humans. An easy-to-use rapid gene amplification method that can directly detect RNA and DNA viruses in clinical samples was developed. This method is based on combining loop-mediated isothermal amplification (LAMP) or reverse transcription-LAMP (RT-LAMP) and RNA GEM Tissue, a thermophilic enzyme that extracts nucleic acid by quickly digesting proteins and ribonucleases. The authors named these methods GEM LAMP and GEM RT-LAMP. These methods were able to detect viral DNA and RNA within 70 min in a single tube using only a water bath. The detection capacities were 10-100-fold more sensitive than those of previously established LAMP and RT-LAMP methods. The GEM LAMP and GEM RT-LAMP methods were used to detect macroscopically the presence of DNA and RNA viruses in sera or fecal samples from cattle, pigs, horses, dolphins, penguins, and sea lions using SYBR green I. The GEM LAMP and GEM RT-LAMP methods thus have considerable versatility as tools for detecting pathogens and are applicable to basic human and veterinary medicine, environmental hygiene, and point-of-care-testing. (C) 2014 Elsevier B.V. All rights reserved.