Determination of the Entomopoxviruses Viral Titer by Quantitative Real Time PCR (qPCR)

Poxviruses are large double-stranded DNA (dsDNA) viruses that infects both human, animals and insects. Most known member of this viruses is Variola virus that caused pox disease and eradicated by vaccination. While other members still infects their hosts it is important to detect, isolate and fight with this viruses. For this purpose determining the titers of viruses is important. Virus titers are used for vaccine development, antiviral activity, genetic engineering, gene therapy and disease treatment studies. Despite all these uses, the methods used to determine virus titer are tedious and time-consuming. We developed a method using quantitative real-time PCR to improve the speed and efficiency of titer determination. In this method, we used a known single-copy gene from AMEV virus, cloned it into a plasmid and generated standard graphs. We measured the virus whose titer was unknown using the standard graph. As a result, a method was developed that can be used to determine the titer of Poxviruses in a simple, fast and effective way.