Applicability of one tube RT-PCR for the detection of bovine viral diarrhea virus (BVDV) in routine laboratory diagnosis

Two commercially available viral RNA extraction kits were tested for their applicability to RT-PCR diagnosis of BVDV infections in comparison with a C atrimox-14(TM) phenol-chloroform RNA extraction method. PCR was carried out as a one tube RT-PCR by application of the Titan(TM) One Tube RT-PCR Syste m (Roche). The combination of the QIAamp Viral RNA Extraction Kit (Qiagen) or the High Pure(TM) Viral RNA Extraction Kit (Roche) with the Titan(TM) On e Tube RT-PCR System turned out to simplify and accelerate the procedure co nsiderably. Serum, plasma, EDTA blood, various tissues, nasal and conjuncti val swabs proved to be suitable specimens for a specific and sensitive RT-P CR detection of BVDV. The RT-PCR yielded higher sensitivity than an antigen ELISA. Sixty-nine pestivirus positive samples which had been detected duri ng this investigation were tested with nested PCR for BVDV type II. No BVDV type It was found within these samples.