二重荧光RT-LAMP方法鉴别检测口蹄疫病毒和水泡性口炎病毒

doi:10.11843/j.issn.0366-6964.2018.05.014

Abstract

Abstract:

Foot-and-mouth disease and vesicular stomatitis are serious cute disease common in cattle, and prevalence in large parts of the word. The aim of this study was to establish a duplex fluorescence-based reverse transcript loop-mediated isothermal amplification assay for detection of foot-and-mouth disease virus (FMDV) and vesicular stomatitis virus (VSV). Two sets of specific primers were designed according to the conserved sequences of FMDV 3D gene and VSV N gene, and the inner primers were synthesized with two different fluorophores at its 5'ends. A duplex fluorescence-based reverse transcript loop-mediated isothermal amplification assay was developed and used for detection of FMDV and VSV. The results showed that this assay could detect at least 100 copies of premixed plasmid containing the targets gene of FMDV and VSV per tube. The specificity of this assay was high, and be able to detect FMDV and VSV without any cross-reactions with other known non-targeted bovine pathogens. Different concentration template of FMDV and VSV could be identified when mixed together without any interference. These findings indicate that this duplex RT-LAMP assay is a simple, rapid, specific, and sensitive method for detection of FMDV and VSV, and can be applied in clinical detection and epidemiological investigation of FMDV and VSV.

CLC Number:

FAN Qing, XIE Zhi-xun, XIE Zhi-qin, XIE Li-ji, HUANG Jiao-ling, ZHANG Yan-fang, ZENG Ting-ting, WANG Sheng, LUO Si-si, DENG Xian-wen, LIU Jia-bo. Development of a Duplex Fluorescence-based Reverse Transcript Loop-mediated Isothermal Amplification Assay for Detection of Foot-and-mouth Disease Virus and Vesicular Stomatitis Virus[J]. ACTA VETERINARIA ET ZOOTECHNICA SINICA, 2018, 49(5): 996-1004.

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Development of a Duplex Fluorescence-based Reverse Transcript Loop-mediated Isothermal Amplification Assay for Detection of Foot-and-mouth Disease Virus and Vesicular Stomatitis Virus

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