Characterisation of the canine parvovirus type 2 variants using minor groove binder probe technology

J Virol Methods. 2006 Apr;133(1):92-9. doi: 10.1016/j.jviromet.2005.10.026. Epub 2005 Nov 28.

Abstract

Two minor groove binder (MGB) probe assays were developed for rapid identification and quantitation of the canine parvovirus type 2 (CPV-2) variants in the faecal samples of dogs. The MGB probe assays using type-specific probes labeled with different fluorophores (FAM and VIC) were able to detect the single nucleotide polymorphisms existing between types 2a/2b and 2b/2c. Both the MGB probe assays were found to be highly specific, sensitive and reproducible, ensuring a precise quantitation of the CPV-2 DNA, as confirmed by comparing the obtained viral DNA loads to those calculated using a real-time TaqMan-based PCR assay established previously. A total of 414 faecal specimens collected from diarrhoeic dogs and tested positive to CPV-2 were analysed by the MGB probe assays and by conventional methods (haemagglutination inhibition with MAbs, PCR-RFLP and sequence analysis) for prediction of the CPV-2 antigen specificity, with an overall agreement of 100%. These results make the established MGB probe assays an attractive tool for direct and rapid characterisation of type 2a, 2b and 2c CPVs.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acid Substitution
  • Animals
  • Antibodies, Monoclonal / metabolism
  • DNA Primers
  • DNA Probes*
  • DNA, Viral / analysis*
  • DNA, Viral / genetics
  • Dog Diseases / genetics
  • Dog Diseases / virology*
  • Dogs
  • Feces / virology
  • Gene Dosage
  • Genetic Variation*
  • Hemagglutination Inhibition Tests
  • Parvoviridae Infections / veterinary*
  • Parvoviridae Infections / virology
  • Parvovirus, Canine / genetics*
  • Parvovirus, Canine / isolation & purification
  • Polymerase Chain Reaction / veterinary
  • Polymorphism, Restriction Fragment Length
  • Polymorphism, Single Nucleotide
  • Taq Polymerase

Substances

  • Antibodies, Monoclonal
  • DNA Primers
  • DNA Probes
  • DNA, Viral
  • Taq Polymerase