Development of a reverse transcription loop-mediated isothermal amplification assay for detection of Porcine teschovirus

J Vet Diagn Invest. 2011 May;23(3):516-8. doi: 10.1177/1040638711403427.

Abstract

Loop-mediated isothermal amplification (LAMP) is a sensitive method for DNA amplification. In the present report, the development of a single-tube, one-step, real-time accelerated reverse transcription (RT)-LAMP for the detection of Porcine teschovirus (PTV) is described. Six designed primers amplified target gene sequences successfully at constant temperature (65 °C) within 1 hr, and the amplification results could be visualized directly by the naked eye. The sensitivity of the LAMP was 10 times higher than that of conventional polymerase chain reaction, and no cross-reactivity was found when the genomes of other common swine pathogens were subjected to the RT-LAMP system. When 43 clinical samples were tested by the RT-LAMP method, results indicated that the test is simple, rapid, accurate, and sensitive for the detection of PTV.

MeSH terms

  • Animals
  • Nucleic Acid Amplification Techniques / methods
  • Nucleic Acid Amplification Techniques / veterinary*
  • Picornaviridae Infections / diagnosis
  • Picornaviridae Infections / veterinary*
  • Picornaviridae Infections / virology
  • Reproducibility of Results
  • Reverse Transcriptase Polymerase Chain Reaction / veterinary
  • Reverse Transcription / genetics
  • Sensitivity and Specificity
  • Swine / virology
  • Swine Diseases / diagnosis
  • Swine Diseases / virology*
  • Teschovirus / genetics*