Development of multiplex polymerase chain reaction for detection of Ehrlichia canis, Babesia spp and Hepatozoon canis in canine blood

Southeast Asian J Trop Med Public Health. 2009 Jan;40(1):35-9.

Abstract

A multiplex polymerase chain reaction (PCR) has been developed for simultaneous detection of canine blood parasites, Ehrlichia canis, Babesia spp and Hepatozoon canis, from blood samples in a single reaction. The multiplex PCR primers were specific to E. canis VirB9, Babesia spp 16S rRNA and H. canis 16S rRNA genes. Specificity of the amplicons was confirmed by DNA sequencing. The assay was evaluated using normal canine and infected blood samples, which were detected by microscopic examination. This multiplex PCR offers scope for simultaneous detection of three important canine blood parasites and should be valuable in monitoring parasite infections in dogs and ticks.

MeSH terms

  • Animals
  • Apicomplexa / genetics*
  • Apicomplexa / isolation & purification
  • Babesia / classification
  • Babesia / genetics*
  • Babesia / isolation & purification
  • Babesiosis / diagnosis
  • Babesiosis / genetics
  • Babesiosis / parasitology
  • Babesiosis / veterinary*
  • Base Sequence
  • DNA Primers / genetics
  • DNA, Protozoan / blood
  • DNA, Protozoan / genetics
  • Dog Diseases / diagnosis*
  • Dog Diseases / genetics
  • Dog Diseases / parasitology
  • Dogs
  • Ehrlichia canis / genetics*
  • Ehrlichia canis / isolation & purification
  • Ehrlichiosis / diagnosis
  • Ehrlichiosis / genetics
  • Ehrlichiosis / microbiology
  • Ehrlichiosis / veterinary*
  • Genes, Bacterial
  • Molecular Sequence Data
  • Polymerase Chain Reaction / methods
  • Polymerase Chain Reaction / veterinary*
  • Polymorphism, Restriction Fragment Length
  • RNA, Ribosomal, 16S / genetics
  • Sensitivity and Specificity
  • Sequence Analysis, DNA

Substances

  • DNA Primers
  • DNA, Protozoan
  • RNA, Ribosomal, 16S