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. 2004 Aug;70(8):4538-43.
doi: 10.1128/AEM.70.8.4538-4543.2004.

Inactivation of caliciviruses

Erwin Duizer  1 , Paul Bijkerk, Barry Rockx, Astrid De Groot, Fleur Twisk, Marion Koopmans
Affiliations

Inactivation of caliciviruses

Erwin Duizer et al. Appl Environ Microbiol. 2004 Aug.

Abstract

The viruses most commonly associated with food- and waterborne outbreaks of gastroenteritis are the noroviruses. The lack of a culture method for noroviruses warrants the use of cultivable model viruses to gain more insight on their transmission routes and inactivation methods. We studied the inactivation of the reported enteric canine calicivirus no. 48 (CaCV) and the respiratory feline calicivirus F9 (FeCV) and correlated inactivation to reduction in PCR units of FeCV, CaCV, and a norovirus. Inactivation of suspended viruses was temperature and time dependent in the range from 0 to 100 degrees C. UV-B radiation from 0 to 150 mJ/cm(2) caused dose-dependent inactivation, with a 3 D (D = 1 log(10)) reduction in infectivity at 34 mJ/cm(2) for both viruses. Inactivation by 70% ethanol was inefficient, with only 3 D reduction after 30 min. Sodium hypochlorite solutions were only effective at >300 ppm. FeCV showed a higher stability at pH <3 and pH >7 than CaCV. For all treatments, detection of viral RNA underestimated the reduction in viral infectivity. Norovirus was never more sensitive than the animal caliciviruses and profoundly more resistant to low and high pH. Overall, both animal viruses showed similar inactivation profiles when exposed to heat or UV-B radiation or when incubated in ethanol or hypochlorite. The low stability of CaCV at low pH suggests that this is not a typical enteric (calici-) virus. The incomplete inactivation by ethanol and the high hypochlorite concentration needed for sufficient virus inactivation point to a concern for decontamination of fomites and surfaces contaminated with noroviruses and virus-safe water.

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Figures

FIG. 1.
FIG. 1.
Times required at different temperatures to achieve a 3 log10 reduction in viable virus titer (TCID50) in DMEM (3 to 4 μg of protein/ml).
FIG. 2.
FIG. 2.
Inactivation of caliciviruses in suspension by UV-B irradiation at 0°C. The dose is presented as cumulative irradiation at 0.43 mJ cm−2 s−1 for different exposure times. Error bars indicate 1 standard deviation (n = 4).
FIG. 3.
FIG. 3.
Inactivation of caliciviruses in suspension incubated for 30 min at 37°C at different pHs. Error bars indicate 1 standard deviation (n = 4).
FIG. 4.
FIG. 4.
Inactivation of caliciviruses in suspension at room temperature by 70% ethanol. Error bars indicate 1 standard deviation (n = 4).
FIG. 5.
FIG. 5.
Inactivation of caliciviruses by 10 min in suspension at room temperature with sodium hypochlorite. Error bars indicate 1 standard deviation (n = 4).
FIG. 6.
FIG. 6.
Ct values for calicivirus and norovirus (NoV) RNA in real-time PCR. RNA was extracted from the stock and diluted before the reverse transcription step. Each data point represents the average Ct ± 1 standard deviation (two experiments, n = 2).
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