Veterinarski arhiv, Vol. 80 No. 1, 2010.
Izvorni znanstveni članak
Molecular detection of Newcastle disease virus using Flinders Tehnology Associates-PCR
Madaboosi S. Narayanan
; Department of Animal Biotechnology, Madras Veterinary, College, Tamil Nadu, India
Manoharan Parthiban
; Department of Animal Biotechnology, Madras Veterinary, College, Tamil Nadu, India
Padmanabhan Sathiya
; Department of Animal Biotechnology, Madras Veterinary, College, Tamil Nadu, India
Kathaperumal Kumanan
; Department of Animal Biotechnology, Madras Veterinary, College, Tamil Nadu, India
Sažetak
The feasibility of using Flinders Technology Associates (FTA) filter papers to store the Newcastle disease virus (NDV), infected allantoic fluid (AF) and tissue samples, for the molecular detection of NDV by reverse transcriptase - polymerase chain reaction (RT-PCR) - was investigated. An FTA card is a cotton based cellulose membrane, with lyophilized chemicals that lyse the viruses and bacteria. The viral RNA was detectable from
FTA cards up to a concentration of 107.6 EID50/100 μL (a 100 times dilution of 109.6 EID50/100 μL of initial stock). The inactivated virus remained stable on the cards for up to 30 days, both at room temperature and 4 ºC. NDV was detected by RT-PCR from all the FTA imprints of the caecal tonsils, kidney, proventriculus, spleen, trachea, faecal swabs and intestinal lesions of NDV-suspected birds. NDV was inactivated upon contact with FTA, as shown by the inability of the virus to propagate in embryonated eggs and its inability to infect chicken embryo fibroblast culture. In conclusion, FTA cards are suitable for collecting and transporting NDV infected samples, without cold storage. The virus inactivated in FTA cards, however, is a suitable source of viral RNA for molecular detection and characterization.
Ključne riječi
Newcastle disease virus; filter paper; molecular detection
Hrčak ID:
53098
URI
Datum izdavanja:
8.3.2010.
Posjeta: 2.969 *