Oxford Expression Technologies - baculoQUANT
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Information on the Oxford Expression Technologies baculoQUANT™ Virus Titration Product
Go to Oxford Expression Technologies website baculoQUANT™ is a novel one-step virus titration kit which can give an accurate result within 2 hours without the need for plaque assays or immunoassays. The baculoQUANT™ One-Step Virus Titration Kit is the quickest method for the titration of baculoviruses, being designed to give accurate titres within 2 - 6 hours (depending on number of viruses). Other methods typically require 24 to 48 hours (immunological assays) or 3 to 4 days (plaque assay or end-point dilution assays). When used to its full capacity of 26 virus titrations, the kit provides very cost effective virus titration assay.
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Literature
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Product Technical Overview
baculoQUANT™
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 baculoQUANT™ is a fast and cost-effective method of recombinant virus titration. It is based on a technique called quantitative PCR (qPCR).  qPCR utilises a set of primers and a probe which are designed to specifically target and amplify a small region of the baculovirus genome. The increase in amplification of this small region is then correlated to virus genome copy number and can be used to measure the number of recombinant virus particles present in an infected insect cell culture.  This novel titration method has been proven to be a very accurate, reproducible and fast way to titrate recombinant baculoviruses, both for single-use titrations and for high throughput applications on automated systems.  

Compared with other methods of virus titration such as plaque assay, end-point dilution and ELISA, which require anywhere from 24 to 120 hours to yield a result and involve specialised procedures, baculoQUANT™ can provide a result within 2 hours, dramatically improving the productivity of the protein production process

Detection is based on quantitative real-time PCR (qPCR) and Taqman fluorogenic probes, specifically designed to the viral genome. Taqman technology takes advantage of the 5 exonuclease activity of Taq polymerase to digest a probe, hybridised between flanking PCR primers, and labeled with two fluorescent dyes. The dyes undergo fluorescent resonance energy transfer (FRET) because of their proximity to each other. Cleavage by Taq during primer elongation interrupts the FRET and allows the reporter dye to fluoresce for every cycle of the PCR. This increase in fluorescence can be analysed in real-time and allows quantification during the exponential phase of the reaction. The system has been calibrated against known viral titres, previously determined by plaque assay.

Each kit contains enough forward and reverse primers, probe and standard virus DNA to enable the user to perform up to 5 qPCR runs, with a total maximum number of 26 viruses titrated.

NB. Virus must be no older than 3 months or an accurate titre cannot be achieved.
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