Use of FTA filter paper for the molecular detection of Newcastle disease virus
| Autor: | Iván Alvarado, Pedro Villegas, Linda B. Purvis, Carlos Estevez, Francisco Perozo |
|---|---|
| Rok vydání: | 2006 |
| Předmět: |
animal structures
Caecal tonsil viruses Newcastle Disease Newcastle disease virus Virulence Biology urologic and male genital diseases Newcastle disease Sensitivity and Specificity Virus Food Animals Animals General Immunology and Microbiology Filter paper Reverse Transcriptase Polymerase Chain Reaction Micropore Filters Nucleic acid sequence bacterial infections and mycoses biology.organism_classification Virology Titer embryonic structures RNA Viral Virus Inactivation Animal Science and Zoology RNA extraction Chickens |
| Zdroj: | Avian pathology : journal of the W.V.P.A. 35(2) |
| ISSN: | 0307-9457 |
| Popis: | The feasibility of using Flinders Technology Associates filter papers (FTA cards) to collect allantoic fluid and chicken tissue samples for Newcastle disease virus (NDV) molecular detection was evaluated. Trizol RNA extraction and one-step reverse transcriptase-polymerase chain reaction (RT-PCR) were used. FTA cards allowed NDV identification from allantoic fluid with a titre of 10(5.8) median embryo lethal doses/ml. The inactivated virus remained stable on the cards for 15 days. NDV was detected from FTA imprints of the trachea, lung, caecal tonsil and cloacal faeces of experimentally infected birds. RT-PCR detection from FTA cards was confirmed by homologous frozen-tissue RT-PCR and virus isolation. Direct nucleotide sequence of the amplified F gene allowed prediction of NDV virulence. No virus isolation was possible from the FTA inactivated samples, indicating viral inactivation upon contact. The FTA cards are suitable for collecting and transporting NDV-positive samples, providing a reliable source of RNA for molecular characterization and a hazard-free sample. |
| Databáze: | OpenAIRE |
| Externí odkaz: |
|