Pseudorabies virus latency: a quantitative approach by polymerase chain reaction.
Some data dealing with the establishment of a quantitative PCR assay are presented. The assay is based on the use of an internal standard (mimic) which differs from the target by a deletion of a few base pairs and which is co-amplified with the target DNA. The resulting PCR products are labelled with fluorescent primers and then separated and detected by an automated sequencer. A highly specific and sensitive PCR assay for the envelope glycoprotein gp50 gene has been developed. This assay is highly reproducible with a detection limit of one copy of PRV DNA. Several mimics were then constructed. As a result we can confirm that the strategy that we have chosen is adequate for the quantification of low amounts of virus DNA present in latently infected swine.
Thiery
R
R
Centre National d'Etudes Vétérinaires et Alimentaires, Laboratoire de Biologie Moléculaire, Ploufragan, France.
Boutin
P
P
Arnauld
C
C
Jestin
A
A
D000818:Animals; D001483:Base Sequence; D017931:DNA Primers; D004279:DNA, Viral / Q000032:analysis*; D004587:Electrophoresis, Agar Gel; D011558:Herpesvirus 1, Suid / Q000502:physiology*; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction / Q000379:methods; D011557:Pseudorabies / Q000175:diagnosis*; D013552:Swine; D013553:Swine Diseases / Q000175:diagnosis*; D014759:Viral Envelope Proteins / Q000235:genetics; D017735:Virus Latency / Q000502:physiology*
D017931:DNA Primers; D004279:DNA, Viral; D014759:Viral Envelope Proteins