Rapid specific and visible detection of porcine circovirus type 3 using loop-mediated isothermal amplification (LAMP)

Transbound Emerg Dis. 2018 Jun;65(3):597-601. doi: 10.1111/tbed.12835. Epub 2018 Mar 4.

Abstract

In this study, a rapid and specific assay for the detection of porcine circovirus type 3 (PCV3) was established using loop-mediated isothermal amplification (LAMP). Four primers were specifically designed to amplify PCV3. The LAMP assay was effectively optimized to amplify PCV3 by water bath at 60°C for 60 min. The detection limit was approximately 1 × 101 copy in this LAMP assay. Compared to porcine circovirus type 2 (PCV2), both gE and gD genes of pseudorabies virus (PRV) and porcine parvovirus (PPV), the LAMP assay showed a high specific detection of PCV3. A visible detection method was developed using SYBR Green I to recognize the results rapidly. Based on the detection of 20 clinical tissue samples, the LAMP assay was more practical and convenient than classical PCR due to its simplicity, high sensitivity, rapidity, specificity, visibility and cost efficiency.

Keywords: PCV3; detection; loop-mediated isothermal amplification (LAMP).

MeSH terms

  • Animals
  • Circovirus / classification
  • Circovirus / genetics*
  • Communicable Diseases, Emerging / veterinary
  • Communicable Diseases, Emerging / virology
  • DNA Primers
  • Nucleic Acid Amplification Techniques / methods*
  • Swine
  • Swine Diseases / virology*

Substances

  • DNA Primers

Associated data

  • GENBANK/KY778776
  • GENBANK/KY778777