Evaluation of feline calicivirus inactivation by cell culture and quantitative real-time PCR
2009
Kumashita, Y., Saraya Co. Ltd., Kashiwara, Osaka (Japan). Biochemical Lab. | Harada, Y. | Takamoto, K. | Murata, Y. | Furuta, T. | Nishio, O.
Some commercially available products claim efficacy against norovirus as evaluated by the polymerase chain reaction (PCR) method. Effects of these products against feline calicivirus (FCV), a norovirus surrogate, was assessed by two different methods, cell culture and quantitative reverse transcriptase real-time PCR (QRT-PCR). Tests showed that the results of the two methods did not always match. Focusing on the components of these products, the efficacy of tannic acid, citric acid and citric acid + 50% (w/w) ethanol against FCV was evaluated by cell culture and QRT-PCR. Using the cell culture method, it was shown that tannic acid was not effective even at high concentrations. 0.5M citric acid and citric acid at concentrations greater than 0.005M + 50% (w/w) ethanol were highly effective against FCV, reducing the infectious titer to levels below the detection limits. On the other hand, using the QRT-PCR method, FCV gene amplification was hardly observed at high concentrations of tannic acid. Furthermore, while almost 100% gene amplification was observed for citric acid by itself even at high concentrations, for citric acid + 50% (w/ w) ethanol, FCV gene amplification was not detected in the citric acid concentration range between 0.5M-0.005M. These findings indicated that there was no correlation between the results from the cell culture and QRT-PCR methods. Therefore, it was concluded that correct results regarding virucidal efficacy cannot always be achieved when only QRT-PCR assays for gene amplification are performed.
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