Naslov
Real-time RT-PCR assay for rapid and specific detection of classical swine fever virus: Comparison of SYBR Green and TaqMan MGB detection methods using novel MBG probes

Autori
Jamnikar Ciglenečki, Urška ; Grom, Joža ; Toplak, Ivan ; Jemeršić, Lorena ; Barlič-Maganja, Darja

Izvornik
Journal of Virological Methods (0166-0934) 147 (2008), 2; 257-264

Vrsta, podvrsta i kategorija rada
Radovi u časopisima, članak, znanstveni

Ključne riječi
Pestivirus; classical swine fever virus; SYBR Green; TaqMan; real time PCR

Sažetak
Real time PCR ia an accurate, rapid and reliable method that can be used for the detection and also for the quantitation of specific DNA molecules.The basic principle is the recurring measurement of a fluorescent signal, which is propotional to the ammount of applification product. In our trial 2 detection systems were tested for classical swine fever virus (CSFV) detection and for its discrimination from other pestiviruses ; non-specific dsDNA- binding dye SYBR Green and specific fluorogenic TaqMan MGB probes. Real time RT-PCR assays were evaluated for diagnostic sensitivity and specificity by different pestiviral reference and field strains. With both approaches, SYBR Green and TaqMan probes, respectively, all of the CSFV strains isolated on cell culture were detected and also clearly distinguished from other pestiviruses. However, the established one step real time TaqMan RT-PCR assay was shown to be more appropriate for pestivirus quantitation, it reduces the risk of contamination and is less time consuming.

Izvorni jezik
Engleski

Znanstvena područja
Veterinarska medicina

POVEZANOST RADA