A quantitative technique for the study of the latency of Aujeszky virus.
Latency represents a challenge for any herpesvirus vaccine. Vaccines could differ in their ability to minimize latency of pseudorabies virus (PRV). To study this possibility, a quantitative and differential PRV-specific polymerase chain reaction (PCR) was developed by the authors. Efficiency of amplification in different tubes is measured by co-amplification with the viral targets (either gI or gp50 genes) of the porcine gene nuclear factor 1. The criteria used to select this approach to a quantitative PCR, as well as for the selection of the standard, are discussed. The amplified products are measured by Southern blot or, alternatively, high performance liquid chromatography. Sensitivity and reproducibility of the technique are evaluated. The ability of this technique to discriminate between the level of latency established by two different PRV strains in trigeminal ganglia is also evaluated. Using this technique, the authors are currently studying whether different vaccines could possess differing levels of ability to minimize, by interference, establishment of wild-type latency.
Department of Veterinary and Biomedical Sciences, University of Nebraska-Lincoln 68583-0905.
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LM
Schang
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Osorio
D000818:Animals; D001483:Base Sequence; D015139:Blotting, Southern / Q000662:veterinary; D002851:Chromatography, High Pressure Liquid / Q000662:veterinary; D017931:DNA Primers / Q000737:chemistry; D015342:DNA Probes / Q000737:chemistry; D004279:DNA, Viral / Q000032:analysis* / Q000737:chemistry; D011558:Herpesvirus 1, Suid / Q000235:genetics / Q000302:isolation & purification / Q000502:physiology*; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction / Q000662:veterinary; D011557:Pseudorabies / Q000276:immunology / Q000382:microbiology*; D011897:Random Allocation; D012008:Recurrence; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D013552:Swine; D013553:Swine Diseases / Q000276:immunology / Q000382:microbiology*; D012668:Trigeminal Ganglion / Q000382:microbiology*
D017931:DNA Primers; D015342:DNA Probes; D004279:DNA, Viral