Sensitive detection of Porcine circovirus-2 by droplet digital polymerase chain reaction

J Vet Diagn Invest. 2015 Nov;27(6):784-8. doi: 10.1177/1040638715608358. Epub 2015 Sep 21.

Abstract

Sensitive detection of Porcine circovirus-2 (PCV-2) is very important for surveillance of postweaning multisystemic wasting syndrome. Droplet digital polymerase chain reaction (ddPCR) is novel PCR method that can achieve high precision. Our study aimed to develop a sensitive assay utilizing ddPCR to detect PCV-2. Specificity of the assay was confirmed by the failure of amplification of DNA of other relevant viruses. The detection limit for ddPCR was 25 copies/μL, a 4-fold greater sensitivity than TaqMan real-time PCR. Both methods showed a high degree of linearity (R(2) = ~1), although TaqMan real-time PCR showed less sensitivity than ddPCR for clinical detection. Our findings indicate that ddPCR might represent a promising platform for detecting PCV-2 viral loads.

Keywords: Detection; Porcine circovirus-2; TaqMan real-time PCR; droplet digital PCR.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • China
  • Circoviridae Infections / diagnosis
  • Circoviridae Infections / veterinary*
  • Circoviridae Infections / virology
  • Circovirus / genetics
  • Circovirus / isolation & purification*
  • DNA, Viral / analysis
  • Limit of Detection
  • Real-Time Polymerase Chain Reaction / methods
  • Real-Time Polymerase Chain Reaction / veterinary*
  • Sensitivity and Specificity
  • Swine
  • Swine Diseases / diagnosis*
  • Swine Diseases / virology
  • Viral Load / veterinary

Substances

  • DNA, Viral