Rapid detection of porcine circovirus type 2 by loop-mediated isothermal amplification

J Virol Methods. 2008 May;149(2):264-8. doi: 10.1016/j.jviromet.2008.01.023. Epub 2008 Mar 19.

Abstract

A method of loop-mediated isothermal amplification (LAMP) was employed to develop a rapid and simple detection system for porcine circovirus type 2 (PCV2). The amplification could be finished in 60 min under isothermal condition at 64 degrees C by employing a set of four primers targeting the cap gene of PCV2. The LAMP assay showed higher sensitivity than the conventional PCR, with a detection limit of five copies per tube of purified PCV2 genomic DNA. No cross-reactivity was observed from the samples of other related viruses including porcine circovirus type 1 (PCV1), porcine parvovirus (PPV), porcine pseudorabies virus (PRV) and porcine reproductive and respiratory syndrome virus (PRRSV). The detection rate of PCV2 LAMP for 86 clinical samples was 96.5% and appeared greater than that of the PCR method. The LAMP assay reported can provide a rapid yet simple test of PCV2 suitable for laboratory diagnosis and pen-side detection due to ease of operation and the requirement of only a regular water bath or heat block for the reaction.

Publication types

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

MeSH terms

  • Animals
  • Circoviridae Infections / diagnosis*
  • Circovirus / genetics
  • Circovirus / isolation & purification*
  • DNA Primers / genetics
  • Nucleic Acid Amplification Techniques / methods*
  • Sensitivity and Specificity
  • Swine
  • Swine Diseases / diagnosis*
  • Temperature
  • Time Factors

Substances

  • DNA Primers