Establishment and application of visual RT-LAMP technology for detecting bovine ephemeral fever virus
Author:
Affiliation:

Clc Number:

Fund Project:

  • Article
  • |
  • Figures
  • |
  • Metrics
  • |
  • Reference
  • |
  • Related
  • |
  • Cited by
  • |
  • Materials
  • |
  • Comments
    Abstract:

    【Objective】To develop a rapid, convenient and visual molecular assay for detecting bovine ephemeral fever virus (BEFV). 【Method】Two pairs of primers based on six conserved regions in G protein gene of BEFV were designed, and the visual reverse transcription loop-mediated isothermal amplification (RT-LAMP) assay for detecting BEFV was developed. The reaction conditions of RT-LAMP assay were optimized, and the assay was compared with PCR.【Result】 The reaction ladder bands from the RTLAMP assay were most obvious when the RT-LAMP reaction system contained 3 mmol·L-1 Mg2+,0.4 mol·L-1 betaine and 1.2 μmol·L-1 dNTPs mix, the ratio of inner to outer primers was 8∶1, and the reaction temperature was 63 ℃. Clear reaction ladder bands were observed after 40 min of reaction. The established RT-LAMP assay had excellent specificity with only BEFV being amplified. The sensitivity was 10 times higher than that of ordinary PCR.【Conclusion】The visual RT-LAMP assay is easy to operate and highly specific, and the results can be conveniently determined. This method can be used for the rapid detection of bovine epidemic fever.

    Reference
    Related
    Cited by
Get Citation

JIA Kun, HAN Taiguang,, YUAN Liguo, SUN Lingshuang, NING Zhangyong, WANG Heng, LI Shoujun. Establishment and application of visual RT-LAMP technology for detecting bovine ephemeral fever virus[J]. Journal of South China Agricultural University,2017,38(3):15-20

Copy
Share
Article Metrics
  • Abstract:
  • PDF:
  • HTML:
  • Cited by:
History
  • Received:June 27,2016
  • Revised:
  • Adopted:
  • Online: May 04,2017
  • Published: