Detection of foot-and-mouth disease virus rna by reverse transcription loop-mediated isothermal amplification

Hao-Tai Chen,Jie Zhang,Xiang-Tao Liu,Yong-Sheng Liu

doi:10.1186/1743-422x-8-510

Abstract

A reverse transcription loop-mediated isothermal amplification (RT-LAMP) assay was developed for foot-and-mouth disease virus (FMDV) RNA. The amplification was able to finish in 45 min under isothermal condition at 64°C by employing a set of four primers targeting FMDV 2B. The assay showed higher sensitivity than RT-PCR. No cross reactivity was observed from other RNA viruses including classical swine fever virus, swine vesicular disease, porcine reproductive and respiratory syndrome virus, Japanese encephalitis virus. Furthermore, the assay correctly detected 84 FMDV positive samples but not 65 FMDV negative specimens. The result indicated the potential usefulness of the technique as a simple and rapid procedure for the detection of FMDV infection.

Highlights

Foot-and-mouth disease virus (FMDV) is a member of the genus Aphthovirus of the family Picornaviridae, which is divided into seven serotypes with no cross-protection conferred among the serotypes [1]
We evaluated the potential of reverse transcription loop-mediated isothermal amplification (RT-LAMP) for the development of a simple and rapid detection system for FMDV RNA
The result indicated that the detection limit of FMDV RT-LAMP was 10 copies whereas that of reverse transcriptase polymerase chain reaction (RT-PCR) was 100 copies per reaction

Summary

Introduction

Foot-and-mouth disease virus (FMDV) is a member of the genus Aphthovirus of the family Picornaviridae, which is divided into seven serotypes with no cross-protection conferred among the serotypes [1]. Due to the aggressive nature of foot-and-mouth disease (FMD), outbreaks usually result in severe economic losses and impact on both national and international trade within the livestock and animal products [2]. Rapid and accurate diagnosis of any suspected FMD cases is of utmost urgency to control this veterinary infection given the extreme contagiousness of the causative virus. Conventional laboratory diagnosis of FMD was made by ELISA detection of specific viral antigens and by observation of cytopathic effects in cell culture [3].

Methods

Results

Conclusion