[Detection and species-specific differentiation of pestiviruses using real-time RT-PCR].
An important prerequisite for an efficient eradication of pestiviruses like bovine viral diarrhea virus or classical swine fever virus are sensitive and specific detection methods. Beside antigen detection with antigen capture ELISAs and virus isolation using cell culture, the detection of virus genomes by reverse transcriptase polymerase chain reaction (RT-PCR) becomes more and more important. By using real-time RT-PCR, the disadvantages of conventional PCR methods concerning the risk of contamination and missing specificity controls are minimized. In the following, the validation and application of different real-time RT-PCR systems for the detection of pestiviruses are presented. In addition, a new "panpesti" probe was tested, and the ability to differentiate between the major species of pestiviruses with known PCR protocols was determined using the Light Cycler PCR machine.
Gaede
Wolfgang
W
Landesamt für Verbraucherschutz Sachsen-Anhalt, Fachbereich Veterinärmedizin, Stendal. wolfgang.gaede@sdl.lav.ms.lsa-net.de
Reiting
Ralf
R
Schirrmeier
Horst
H
Depner
Klaus Robert
KR
Beer
Martin
M
D000818:Animals; D004279:DNA, Viral / Q000032:analysis; D010576:Pestivirus / Q000145:classification* / Q000235:genetics / Q000302:isolation & purification*; D018182:Pestivirus Infections / Q000175:diagnosis / Q000662:veterinary*; D010802:Phylogeny; D015203:Reproducibility of Results; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000379:methods / Q000662:veterinary*; D012680:Sensitivity and Specificity; D013045:Species Specificity; D013997:Time Factors
D004279:DNA, Viral