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Title: USE OF FTA FILTER PAPER FOR THE MOLECULAR DETECTION OF NEWCASTLE DISEASE VIRUS

Author
item PEROZO, FRANCISCO - UNIV OF GEORGIA-ATHENS,GA
item VILLEGAS, PEDRO - UNIV OF GEORGIA-ATHENS,GA
item Estevez, Carlos
item ALVARADO, IVAN - UNIV OF GEORGIA-ATHENS,GA
item PURVIS, LINDA - UNIV OF GEORGIA-ATHENS

Submitted to: Avian Pathology
Publication Type: Peer Reviewed Journal
Publication Acceptance Date: 10/10/2005
Publication Date: 4/15/2006
Citation: Perozo, F., Villegas, P., Estevez, C., Alvarado, I., Purvis, L.B. 2006. Use of FTA filter paper for the molecular detection of Newcastle disease virus. Avian Pathology. 35(2):93-98.

Interpretive Summary: The research work relates to the use of a paper card impregnated with chemical compounds capable of inactivating biological samples for their safe transport. These cards can inactivate viruses and bacteria, and still preserve the integrity of the organisms’ nucleic acids, which can later be used for molecular diagnostic procedures. In the present work, the effectiveness for sample inactivation and ability to preserve nucleic acids from pathogens of these cards (FTA cards) have been determined by the use of said cards in samples obtained from Newcastle disease virus-infected embryonating chicken eggs’ allantoic fluid and tissues from virus-infected chickens.

Technical Abstract: The feasibility of using FTA® cards to collect allantoic fluid (AF) and chicken tissue samples for Newcastle diseases 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 AF with titre of 105.8 ELD50/ml. The inactivated virus remained stable on the cards for 15 days. NDV was detected from FTA® imprints of 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.