An efficient recovery method for enteric viral particles from agricultural soils

Citation

Brassard, J., Gagné, M.J. (2018). An efficient recovery method for enteric viral particles from agricultural soils. Journal of Virological Methods, [online] 261 1-5. http://dx.doi.org/10.1016/j.jviromet.2018.06.014

Plain language summary

Enteric viruses are recognized as the leading cause of epidemics of gastroenteritis and hepatitis worldwide. A better understanding of their prevalence and persistence in the environment is needed to develop effective control strategies. The aim of this study was to develop an efficient procedure for recovering infectious viral particles from agricultural soils. Samples (25 g) of soil (black soil, silty soil and sandy soil) were inoculated with murine norovirus (MNV) and feline calicivirus (FCV) viruses and mixed with five different buffers, and the viral genetic material was extracted using three commercial kits. The combination of the modified Eagle’s buffer solution followed by the magnetic bead-based nucleic acid extraction kit was identified as the most effective procedure for preserving the infectivity of viral particles and removing inhibitors. In addition, the results presented in this study highlight the importance of using an internal process control, such as artificial inoculation with MNV at known concentrations during detection by molecular methods, in order to avoid the occurrence of false negative reactions

Abstract

Enteric viruses have been recognized as the leading cause of non-bacterial gastroenteritis and hepatitis outbreaks around the world. Understanding their prevalence and persistence in the environment is important for the effective control of these infections. The aim of this study was to develop an efficient recovery procedure for viral infectious particles from agricultural soils. Samples (25 g) of soil (black earth soil, loamy soil, and sandy soil) were spiked with murine norovirus (MNV) and feline calicivirus (FCV), mixed with five different buffers and viral genetic material was extracted by 3 commercial kits. The combination consisted by the modified Eagle's medium buffer followed by Dynabeads nucleic acid extraction kit, when the detection is conducted by molecular biology, has been identified as being the most effective procedure to preserve the viral particle infectivity and also to remove PCR inhibitors. The recovery percentages of infectious MNV for the 3 types of soils were 54.3%, 54.4%, and 56.9%. In contrast, the titres of the FCV varied depending on the type of soil, and the recovery percentages were 47.8% in the black soil, 15.6% in the loamy soil, and 17.7% in the sandy soil. Also, the results presented in this study highlight the importance of using an internal process control such as artificial inoculation with MNV at known concentrations during detection by molecular methods, in order to avoid the occurrence of false negative reactions.

Publication date

2018-11-01

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