A DNA miniarray system for simultaneous visual detection of porcine circovirus type 1 (PCV1) and 2 (PCV2) in pigs

Vet Res Commun. 2009 Feb;33(2):139-47. doi: 10.1007/s11259-008-9080-8. Epub 2008 Jul 24.

Abstract

A miniarray system was developed for the simultaneous detection of porcine circovirus type 1 (PCV1) and type 2 (PCV2) in pigs. The system consists of a polymerase chain reaction (PCR) step to amplify target viral DNA, followed by detection of the amplified DNA using a membrane-anchored probe array and an avidin-alkaline phosphatase (Av-AP) indicator system. The lower limit of detection of PCV using the miniarray was 10(1.9) tissue culture infectious dose 50 (TCID(50))/ml and 10(2.08)TCID(50)/ml for PCV1 and PCV2, respectively, and 100 viral copies/microl for both PCV1 and PCV2. We validated the miniarray system using 141 lymph node specimens from pigs with suspected postweaning multisystemic wasting syndrome or porcine dermatitis and nephropathy syndrome. Of the 141 samples evaluated, 55 were identified as positive for PCV by the miniarray. Relative to in situ hybridization, the sensitivity and specificity of the miniarray was 100% and 98.9%, respectively. In contrast to other microarray systems, the miniarray does not require a DNA chip reader, since the results can be determined by visual inspection of colorized spots on a nylon membrane. This system represents an effective alternative method for the differential detection of PCV1 and PCV2 in pigs, as well as the maintenance of PCV-free cell lines and pre-screening of commercial vaccines for possible contamination.

Publication types

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

MeSH terms

  • Animals
  • Circoviridae Infections / diagnosis
  • Circoviridae Infections / veterinary*
  • Circoviridae Infections / virology
  • Circovirus / genetics
  • Circovirus / isolation & purification*
  • DNA, Viral / chemistry
  • DNA, Viral / genetics
  • In Situ Hybridization / veterinary
  • Lymph Nodes / virology
  • Oligonucleotide Array Sequence Analysis / methods
  • Oligonucleotide Array Sequence Analysis / veterinary*
  • Porcine Postweaning Multisystemic Wasting Syndrome / diagnosis
  • Porcine Postweaning Multisystemic Wasting Syndrome / virology*
  • Sensitivity and Specificity
  • Swine

Substances

  • DNA, Viral