Reverse transcription loop-mediated isothermal amplification (RT-LAMP) for rapid detection of viral hemorrhagic septicaemia virus (VHS)

被引：71

作者：

Soliman, H. ^{[1
]}

El-Matbouli, M. ^{[1
]}

机构：

[1] Univ Munich, Fac Med Vet, Inst Zool Fish Biol & Fish Dis, D-80539 Munich, Germany

来源：

VETERINARY MICROBIOLOGY | 2006年 / 114卷 / 3-4期

关键词：

RT-LAMP; VHS; diagnosis; SYBR Green I; RT-PCR;

D O I：

10.1016/j.vetmic.2005.11.063

中图分类号：

Q93 [微生物学];

学科分类号：

071005 ; 100705 ;

摘要：

A one step reverse transcription loop-mediated isothermal amplification (RT-LAMP) assay was developed for detection of viral hemorrhagic septicaemia virus (VHS). A set of six primers were designed, based on the G-protein sequence of the VHS virus serotypes (He, Fl, 23.75, Klapmolle and Rindsholm). The assay was optimised to amplify VHS RNA by incubation at 63 degrees C for only 1 h, and required only a simple water bath or heating block to provide a constant temperature of 63 degrees C. RT-LAMP amplification products were detected by visual inspection using SYBR Green I stain and had a ladder-like appearance when electrophoresed on an agarose gel. The detection limit of the RT-LAMP assay was found to be similar to the commonly used RTPCR method: both methods detected VHS RNA at a dilution of 10(6). The assay was evaluated using clinical samples and the results indicated the suitability and simplicity of the test as a rapid, field diagnostic tool for VHS virus. (c) 2005 Elsevier B.V. All rights reserved.

引用

页码：205 / 213

页数：9

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