Development of Real-Time RT-PCR for the Detection of Avian Influenza Virus
| Autor: | Kenton L. Lohman, Michael L. Perdue, Dennis A. Senne, L. L. Bulaga, Luke T. Daum, Erica Spackman, L. Garber, David L. Suarez, T. J. Myers |
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| Rok vydání: | 2003 |
| Předmět: |
Hemagglutinin (influenza)
Enzyme-Linked Immunosorbent Assay Biology urologic and male genital diseases medicine.disease_cause Sensitivity and Specificity H5N1 genetic structure law.invention Food Animals law medicine Animals Gene Poultry Diseases Polymerase chain reaction Struthioniformes Hemagglutination assay General Immunology and Microbiology Bird Diseases Reverse Transcriptase Polymerase Chain Reaction virus diseases Virology female genital diseases and pregnancy complications Reverse transcriptase Influenza A virus subtype H5N1 Ducks Real-time polymerase chain reaction Influenza A virus Influenza in Birds biology.protein Animal Science and Zoology Chickens |
| Zdroj: | Avian Diseases. 47:1079-1082 |
| ISSN: | 1938-4351 0005-2086 |
| DOI: | 10.1637/0005-2086-47.s3.1079 |
| Popis: | A real-time reverse transcriptase/polymerase chain reaction (RRT-PCR) assay was developed using hydrolysis probes for the detection of avian influenza virus (AIV) and the H5 and H7 subtypes. The AIV specific primers and probes were directed to regions of the AIV matrix gene that are conserved among most type A influenza viruses. The H5 and H7 primers and probes are directed to H5 and H7 hemagglutinin gene regions that are conserved among North American avian influenza viruses. The sensitivity and specificity of this RRT-PCR assay was compared to virus isolation (VI) in chicken embryos with 1550 clinical swab samples from 109 live-bird markets (LBMs) in New York and New Jersey. RRT-PCR detected influenza in samples from 61 of 65 (93.8%) of the LBMs that were the sources of VI positive samples. Of the 58 markets that were positive for H7 influenza by hemagglutination inhibition assay, RRT-PCR detected H7 influenza in 56 markets (96.5%). Too few H5 positive samples were obtained to validate the H5 RRT-PCR assay in this study. Although RRT-PCR was less sensitive than VI on an individual sample basis, this study demonstrated that the AIV and H7 RRT-PCR assays are good tools for the rapid screening of flocks and LBMs. |
| Databáze: | OpenAIRE |
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