Detection of Chlamydia psittaci DNA in avian clinical samples by polymerase chain reaction

R. Glyn Hewinson, Peter C. Griffiths, Bob J. Bevan, Sharon E.S. Kirwan, Mavis E. Field, Martin J. Woodward, Michael Dawson*

*Corresponding author for this work

Research output: Contribution to journalArticlepeer-review

59 Citations (SciVal)

Abstract

A polymerase chain reaction (PCR) assay was developed to detect Chlamydia psittaci DNA in faeces and tissue samples from avian species. Primers were designed to amplify a 264 bp product derived from part of the 5' non-translated region and part of the coding region of the ompA gene which encodes the major outer membrane protein. Amplified sequences were confirmed by Southern hybridization using an internal probe. The sensitivity of the combined assay was found to be between 60 to 600 fg of chlamydial DNA (approximately 6 to 60 genome copies). The specificity of the assay was confirmed since PCR product was not obtained from samples containing several serotypes of C. trachomatis, strains of C. pneumoniae, the type strain of C. pecorum, nor from samples containing microorganisms commonly found in the avian gut flora. In this study, 404 avian faeces and 141 avian tissue samples received by the Central Veterinary Laboratory over a 6 month period were analysed by PCR, antigen detection ELISA and where possible, cell culture isolation. PCR performed favourably compared with ELISA and cell culture, or with ELISA alone. The PCR assay was especially suited to the detection of C. psittaci DNA in avian faeces samples. The test was also useful when applied to tissue samples from small contact birds associated with a case of human psittacosis where ELISA results were negative and chlamydial isolation was a less favourable method due to the need for rapid diagnosis.

Original languageEnglish
Pages (from-to)155-166
Number of pages12
JournalVeterinary Microbiology
Volume54
Issue number2
DOIs
Publication statusPublished - 01 Feb 1997

Keywords

  • Chlamydia psittaci
  • polymerase chain reaction
  • Cell Line
  • Genes, Bacterial
  • Bacterial Outer Membrane Proteins/genetics
  • Humans
  • Parrots
  • Bird Diseases
  • DNA Primers
  • Chick Embryo
  • Psittacosis/diagnosis
  • Animals
  • Polymerase Chain Reaction/methods
  • Feces/microbiology
  • Sensitivity and Specificity
  • Enzyme-Linked Immunosorbent Assay/methods
  • Chlamydophila psittaci/genetics
  • DNA, Bacterial/analysis

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