Real-time fluorescence loop-mediated isothermal amplification for the diagnosis of hemorrhagic enteritis virus

Virus Res. 2014 Apr:183:50-5. doi: 10.1016/j.virusres.2014.01.023. Epub 2014 Jan 31.

Abstract

Suspected cases of hemorrhagic enteritis associated with hemorrhagic enteritis virus (HEV) are becoming more frequent among yellow chickens in the Guangdong Province of China. In this study, we have developed a one-step, ecumenical, real-time fluorescence loop-mediated isothermal amplification (RealAmp) assay for the rapid diagnosis of HEV. The RealAmp assay was performed at 63°C and reduced the assay time to 15min, using a simple and portable device, the ESE-Quant Tube Scanner. The detection limit of DNA was 1fg/μl, and the detection was specific only to HEV. We also used nested PCR to evaluate the application of the RealAmp assay. The coincidence rate of the two methods was 100%. Our data indicated that the RealAmp assay provides a sensitive, specific, and user-friendly diagnostic tool for the identification and quantification of HEV for field diagnosis and in laboratory research.

Keywords: Hemorrhagic enteritis virus; Nested PCR; RealAmp.

Publication types

  • Evaluation Study
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Adenoviridae Infections / veterinary*
  • Adenoviridae Infections / virology
  • Animals
  • Chickens
  • China
  • Enteritis / veterinary*
  • Enteritis / virology
  • Molecular Diagnostic Techniques / methods*
  • Nucleic Acid Amplification Techniques / methods*
  • Poultry Diseases / diagnosis*
  • Poultry Diseases / virology
  • Sensitivity and Specificity
  • Siadenovirus / isolation & purification*
  • Temperature
  • Time Factors
  • Veterinary Medicine / methods
  • Virology / methods*