Detection and differentiation of virulent and avirulent strains of Newcastle disease virus by real-time PCR
| Autor: | Lukasz Rabalski, Beata Gromadzka, Dawid Nidzworski |
|---|---|
| Rok vydání: | 2011 |
| Předmět: |
Paramyxoviridae
AIV avian influenza virus APMV avian paramyxovirus Newcastle disease virus Virulence Diamines Polymerase Chain Reaction Sensitivity and Specificity Newcastle disease Article Melting curve analysis Virus law.invention Microbiology chemistry.chemical_compound PCR polymerase chain reaction law Virology Transition Temperature Benzothiazoles CSFV classical swine fever virus Organic Chemicals Rubulavirus Polymerase chain reaction ICPI intercerebral pathogenicity index Staining and Labeling biology SYBR Green I biology.organism_classification Detection chemistry NDV Newcastle disease virus Differentiation Quinolines EID egg infective dose Real-time PCR |
| Zdroj: | Journal of Virological Methods |
| ISSN: | 0166-0934 |
| DOI: | 10.1016/j.jviromet.2010.12.015 |
| Popis: | A rapid diagnostic method based on the melting curve SYBR Green I real-time PCR analysis was developed to detect and differentiate Newcastle disease virus (NDV) strains. Degenerated primers based on the cleavage site sequence of the F0 gene were designed to detect specific sequences characteristic of virulent and avirulent strains of NDV. Eighteen strains of NDV from four lineages were identified and grouped into virulent and avirulent strains. Peaks on the melting temperature graph with melting temperature values between 80.00 and 83.80°C were observed for lentogenic (avirulent) strains. T(m) values higher than 83.80 were observed for virulent (mesogenic and velogenic) strains. The detection limit of real-time PCR was 2 × 10(2) plasmid copies per reaction or 10(2) EID(50) for velogenic strains and 10(3) EID(50) for lentogenic strains. The results obtained in this study demonstrate the possible applications for melting curve real-time PCR analysis in laboratory practice for the diagnosis and differentiation of avirulent and virulent strains of Newcastle disease virus. |
| Databáze: | OpenAIRE |
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