Avtor/Urednik     Barlič-Maganja, Darja; Grom, Jože
Naslov     Molekularna diagnostika pestivirusov
Tip     članek
Vir     In: Bole-Hribovšek V, Ocepek M, Klun N, editors. Zbornik s programom 2. kongres slovenskih mikrobiologov z mednarodno udeležbo; 1998 sep 27-30; Portorož. Ljubljana: Slovensko mikrobiološko društvo,
Leto izdaje     1998
Obseg     str. 408-12
Jezik     slo
Abstrakt     Recent work represents three different methods based on specific amplification of the pestivirus RNA by reverse transcription and polymerase chain reaction (RT-PCR). Two techniques for simultaneous detection and differentiation of bovine viral diarrhoea virus (BVDV) and classical swine fewer virus (CSFV), nested-PCR and PCR-ELISA, have been developed. Primers for pestivirus amplification and BVDV-specific or CSFV-specific nested reamplification were selected from two genome regions. 5'-noncoding region (NCR) is highly conserved and is suitable for primer selection for all pestivirus isolated detection. For pestivirus differentiation gene region coding for Npro, C and E0 protein was selected. Primers for BVDV-specific amplication and specific probes for BVDV and CSFV discrimination were selected from this region. The electrophoretic analysis of the PCR products and detection of the amplification products by strain specific capture probe hybridisation and colorimetric assay in microwell plates were compared. By serial dilutions of PCR products the PCR-ELISA was found to be 100 times more sensitive than the conventional agarose gel electrophoresis. Thus PCR-ELISA offers the possibility to avoid nested-PCR and eventual contamination between samples.
Deskriptorji     PESTIVIRUS INFECTIONS
CATTLE
SWINE
HOG CHOLERA VIRUS
BOVINE VIRUS DIARRHEA-MUCOSAL DISEASE
POLYMERASE CHAIN REACTION
RNA-DIRECTED DNA POLYMERASE
ENZYME-LINKED IMMUNOSORBENT ASSAY