Development of conventional multiplex PCR method for discrimination between Dispharynx nasuta and Cheilospirura hamulosa (Nematoda: Acuariidae) parasitizing poultry

J Vet Med Sci. 2021 Feb 25;83(2):226-229. doi: 10.1292/jvms.20-0664. Epub 2020 Dec 18.

Abstract

The poultry infections caused by Dispharynx nasuta and Cheilospirura hamulosa nematodes are difficult to be diagnosed by fecal examination because of their egg similarity. In this study, we analyzed DNA sequences of nuclear ribosomal 18S-ITS1-5.8S-ITS2-28S region of D. nasuta and C. hamulosa and developed conventional multiplex PCR method using species-specific primers for discriminating between the two species. The method amplified 455-bp and 319-bp fragments specific to D. nasuta and C. hamulosa, respectively, and did not produce them against the other chicken nematode species, Ascaridia galli, Oxyspirura mansoni, Heterakis gallinarum, Heterakis beramporia, and Heterakis indica, suggesting that the multiplex PCR is sensitive and available for species diagnosis.

Keywords: Cheilospirura hamulosa; Dispharynx nasuta; multiplex PCR; nuclear ribosomal 18S-ITS1-5.8S-ITS2-28S DNA; poultry.

MeSH terms

  • Animals
  • Base Sequence
  • Chickens / parasitology*
  • DNA, Ribosomal / chemistry
  • DNA, Ribosomal / isolation & purification
  • Female
  • Male
  • Multiplex Polymerase Chain Reaction / methods
  • Multiplex Polymerase Chain Reaction / veterinary*
  • Poultry
  • Poultry Diseases / diagnosis
  • Poultry Diseases / parasitology*
  • RNA, Ribosomal / genetics
  • Sequence Alignment / veterinary
  • Spirurida Infections / diagnosis
  • Spirurida Infections / parasitology
  • Spirurida Infections / veterinary*
  • Spirurina / classification
  • Spirurina / genetics*

Substances

  • DNA, Ribosomal
  • RNA, Ribosomal