Comparison of two elution buffers used in an adsorption-elution method with a negatively charged membrane to recover Norovirus from lettuce (Lactuca sativa)
DOI:
https://doi.org/10.3395/vd.v2n3.195Keywords:
Norovirus, Filtration, Lettuce, PP7 Bacteriophage, Internal Control ProcessAbstract
Phosphate saline buffer (PBS) and glycine buffer (GB) were evaluated as elution buffers in an adsorption-elution method using a negatively charged membrane associated with quantitative polymerase chain reaction (qPCR) and seminested PCR for detection of Norovirus genogroup II (NoV GII) from lettuce. In this methodology, PP7 bacterio-phage was used as a virus sample for process control. The qPCR showed more sensitivity than semi-nested PCR for NoV GII detection. The recovery efficiency, using PBS and GB, ranged from 24.72 to 60.78% and 19.48 to 137.26% for NoV GII, and from 0.01 to 0.15% and 0.13 to 6.04% for PP7 bacteriophage, respectively. Elution with GB was more effi-cient for PP7 bacteriophage recovery (p = 0.03), but no difference was seen for NoV GII (p = 0.57). The GB performed better than PBS as an eluent solution and can be consid-ered a methodological improvement.
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