Development of reverse-transcription loop-mediated isothermal amplification for the detection of infectious bursal disease virus

J Virol Methods. 2009 Dec;162(1-2):267-71. doi: 10.1016/j.jviromet.2009.07.010. Epub 2009 Jul 28.

Abstract

To establish a reverse-transcription loop-mediated isothermal amplification (RT-LAMP) method for rapid detection of infectious bursal disease virus (IBDV), four primers specific to six regions of the VP3 gene were designed; the VP3 region was selected because it is a conserved part of the IBDV genome. After amplification in an isothermal water bath for 70 min, samples containing IBDV generated the expected ladder-like products while other viruses generated no product. The sensitivity and specificity of the RT-LAMP assay were evaluated by comparison with reverse-transcription polymerase chain reaction (RT-PCR) and virus isolation. The assay was significantly more sensitive than normal gel-based RT-PCR. Because it is specific and simple, the RT-LAMP assay can be widely applied in clinical laboratories for rapid detection of IBVD.

Publication types

  • Evaluation Study

MeSH terms

  • Animals
  • Birnaviridae Infections / diagnosis
  • Birnaviridae Infections / veterinary
  • Birnaviridae Infections / virology
  • Chick Embryo
  • Chickens / virology
  • DNA Primers
  • Infectious bursal disease virus* / genetics
  • Infectious bursal disease virus* / isolation & purification
  • Nucleic Acid Amplification Techniques / methods*
  • Poultry Diseases / diagnosis
  • Poultry Diseases / virology
  • Reverse Transcriptase Polymerase Chain Reaction / methods*
  • Sensitivity and Specificity
  • Sequence Analysis, DNA
  • Viral Structural Proteins / genetics

Substances

  • DNA Primers
  • VP3 protein, infectious bursal disease virus
  • Viral Structural Proteins