Development of methods for detection and quantification of avian influenza and Newcastle disease viruses in compost by real-time reverse transcription polymerase chain reaction and virus isolation
| Autor: | Brian W. Brooks, J. Pasick, J. L. Spencer, B. Ma, C. Grenier, D. C. Wilkie, Maria Chan, Jiewen Guan |
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| Rok vydání: | 2008 |
| Předmět: |
animal structures
animal diseases viruses Newcastle disease virus engineering.material medicine.disease_cause complex mixtures Newcastle disease Virus Microbiology Feces Soil medicine Animals Soil Microbiology biology Compost Reverse Transcriptase Polymerase Chain Reaction fungi Embryonated RNA General Medicine biology.organism_classification Virology Influenza A virus subtype H5N1 Specific Pathogen-Free Organisms Reverse transcription polymerase chain reaction Manure Influenza A virus engineering RNA Viral Animal Science and Zoology Chicken manure Chickens |
| Zdroj: | Poultry science. 87(5) |
| ISSN: | 0032-5791 |
| Popis: | Composting has been used for disposal of poultry carcasses and manure following outbreaks caused by avian influenza virus (AIV) and Newcastle disease virus (NDV), but methods are needed to test for survival of the viruses in compost to ensure biosecurity. Methods developed in the present study include extracting viruses from compost and purifying viral RNA. The extracted viruses were detected by virus isolation using embryonated chicken eggs, and the purified RNA was detected by real-time reverse transcription PCR (RRT-PCR). The virus isolation and the RRT-PCR methods were evaluated with 3 compost preparations that were produced from chicken manure mixed with corn silage, wood shavings, or wheat straw. The detection limits of both methods were 1,700 and 1,000 embryo lethal doses of AIV and NDV per gram of compost, respectively. The copy number of viral RNA quantified by RRT-PCR was highly correlated with the amount of virus in compost. The results suggested that the RRT-PCR method may be used as an alternative to the virus isolation method for rapid detection and accurate quantification of AIV and NDV in compost. |
| Databáze: | OpenAIRE |
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