Detection of Chlamydia psittaci DNA in avian clinical samples by polymerase chain reaction |
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| Institution: | 1. Department of Bacteriology, Veterinary Laboratories Agency, Central Veterinary Laboratory, New Haw, Addlestone, Surrey KT15 3NB, UK;2. Department of Virology, Veterinary Laboratories Agency, Central Veterinary Laboratory, New Haw, Addlestone, Surrey KT15 3NB, UK;1. Novartis Vaccines, Via Fiorentina 1, 53100 Siena, Italy;1. Institute of Immunology, Centre de Recherche Public de la Santé/Laboratoire National de Santé, 20A rue Auguste Lumière, L-1950 Luxembourg, Luxembourg;2. Department of Veterinary Medicine, University of Ibadan, Ibadan, Oyo State, Nigeria;3. District Hospital Bali, North West Regional Delegation of Public Health, Bamenda, Northwest Province, Cameroon;3. Biodesign Institute, Center for Innovations in Medicine, Arizona State University, Tempe, AZ;4. Department of Biochemistry, Geisel School of Medicine at Dartmouth, Hanover, NH;1. Institute of Social and Preventive Medicine, University of Bern, Bern, Switzerland;2. Department of Clinical Veterinary Medicine, Vetsuisse Faculty, University of Bern, Bern, Switzerland;3. Section of Epidemiology, Vetsuisse Faculty, University of Zurich, Zurich, Switzerland;1. University Paris-Est, Anses, Animal Health Laboratory, Bacterial Zoonoses Unit, Maisons-Alfort, France;2. Robert Koch Institute, MF1 Bioinformatics, Berlin, Germany;3. Friedrich-Loeffler-Institut (Federal Research Institute for Animal Health), Institute of Molecular Pathogenesis, Germany;4. Friedrich-Schiller-Universität Jena, RNA Bioinformatics and High-Throughput Analysis, Jena, Germany;5. University Paris-Est, Anses, Food Research Laboratory, IdentyPath Platform, Maisons-Alfort, France;6. Tour du Valat, Centre de recherche pour la conservation des zones humides méditerranéennes, Le Sambuc, Arles, France;7. Bioparc - Zoo de Doué la fontaine, 103 rue de Cholet, 49700 Doué la Fontaine, France;8. Parc Zoologique de Paris, avenue de Daumesnil, 75012 Paris, France;9. Unit of Viral Genetics and Biosafety, ANSES, Laboratory of Ploufragan, Ploufragan, France;10. University of Maryland, Electron Microscopy Core Imaging Facility, Baltimore, MD 21201, USA;11. University of Maryland, Department of Microbial Pathogenesis, Baltimore, MD 21201, USA;12. Marine Microbiology Group, Department of Animal and Microbial Biodiversity, Mediterranean Institute for Advanced Studies, 07190 Esporles, Spain;1. Jiangsu Co-innovation Center for Prevention and Control of Important Animal Infectious Diseases and Zoonoses, Yangzhou University College of Veterinary Medicine, Yangzhou, Jiangsu, 225009, PR China;2. College of Animal Science, Anhui Science and Technology University, Anhui, China;3. College of Veterinary Medicine, Auburn University, Auburn, AL, USA;4. Friedrich-Loeffler-Institut (Federal Research Institute for Animal Health), Jena, Germany;5. Yangzhou University College of Animal Science, Yangzhou, Jiangsu, 225009, PR China |
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| 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. |
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