Author/Editor		Barlič-Maganja, Darja; Grom, Jože
Title		Molekularna diagnostika pestivirusov
Type		članek
Source		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,
Publication year		1998
Volume		str. 408-12
Language		slo
Abstract		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.
Descriptors		PESTIVIRUS INFECTIONS CATTLE SWINE HOG CHOLERA VIRUS BOVINE VIRUS DIARRHEA-MUCOSAL DISEASE POLYMERASE CHAIN REACTION RNA-DIRECTED DNA POLYMERASE ENZYME-LINKED IMMUNOSORBENT ASSAY