Development of probes to differentiate porcine circovirus types 1 and 2 in vitro by in situ hybridization

Porntippa Nawagitgul, Igor Morozov, Theerapol Sirinarumitr, Steven D. Sorden, Prem S. Paul

Research output: Contribution to journalArticle

9 Scopus citations

Abstract

Porcine circovirus type 1 (PCV1), a PK-15 cell line contaminant, and porcine circovirus type 2 (PCV2), associated with post-weaning multisystemic wasting syndrome (PMWS), are genetically and antigenically related. Several techniques have been developed to detect PCV, including in situ hybridization (ISH). Previously reported probes used for ISH may hybridize with both PCV1 and PCV2 nucleic acids. We attempted to produce probes for ISH that can detect and differentiate PCV2 from PCV1 in PCV-infected cells. Riboprobes were synthesized from the sense and antisense strands of both open reading frames 1 and 2 (ORF1 and ORF2) of PCV2. At 42 and 58°C, the ORF1 antisense probe hybridized with nucleic acid from both PCV1- and PCV2-infected cells. At 58°C, the ORF2 antisense probe hybridized with PCV2 nucleic acid but not with PCV1 nucleic acid. The ORF1 and ORF2 sense probes bound only with PCV2 nucleic acid. Both antisense strand probes produced stronger signals than the sense strand probes. The results showed that the PCV2 ORF1 antisense probe is the most likely probe to detect both PCV types while the ORF2 antisense probe is capable of discriminating between PCV1 and PCV2. Copyright (C) 2000 Elsevier Science B.V.

Original languageEnglish (US)
Pages (from-to)83-89
Number of pages7
JournalVeterinary Microbiology
Volume75
Issue number1
DOIs
StatePublished - Jul 3 2000

Keywords

  • In situ hybridization (ISH)
  • Porcine circovirus (PCV)

ASJC Scopus subject areas

  • Microbiology
  • veterinary(all)

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