Duplex shuttle PCR for differential diagnosis of budgerigar fledgling disease and psittacine beak and feather disease

Hirohito Ogawa, Tsuyoshi Yamaguchi, Hideto Fukushi

Research output: Contribution to journalArticle

33 Citations (Scopus)

Abstract

Two common viral diseases in psittacine birds including budgerigar fledgling disease (BFD), generally called avian polyomavirus (APV) infection, and psittacine beak and feather disease (PBFD) have similar clinical manifestations characterized by feather disorders. A duplex shuttle PCR was developed for detection of APV and PBFD virus (PBFDV). Two pairs of oligonucleotide primers were designed to amplify a 298-bp fragment of the t/T antigen region of APV genome and a 495-bp fragment of the capsid protein region encoded by open reading frame (OMF) C1 of PBFDV genome, respectively. Ie the present study, APV and PBFDV were detected simultaneously in one tube by duplex shuttle PCR using these two pairs of primers. The detection limits were 2 viral copies of APV and 3 viral copies of PBFDV. In the clinical application, we detected 16 APV-positive, 15 PBFDV-positive, and 3 mixed infected samples in 39 samples examined. Sequences of the amplified products were read. The t/T antigen region was conserved in the APV-positive samples as expected. ORF C1 of PBFDV genome showed diversity. Phylogenic analysis indicated that PBFDV ORF C1 consisted of 6 clusters which were related to subfamilies of psittacine birds. Our duplex shuttle PCR could be a useful method for differential diagnosis and molecular epidemiology of BFD and PBFD.

Original languageEnglish
Pages (from-to)227-237
Number of pages11
JournalMICROBIOLOGY and IMMUNOLOGY
Volume49
Issue number3
DOIs
Publication statusPublished - 2005

Keywords

  • Avian polyomavirus
  • Budgerigar fledgling disease
  • Duplex PCR
  • Psittacine beak and feather disease

ASJC Scopus subject areas

  • Microbiology
  • Immunology
  • Virology

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