Comparison of two elution buffers used in an adsorption-elution method with a negatively charged membrane to recover Norovirus from lettuce (Lactuca sativa)

• Main Authors: Marcelo Luiz Lima Brandão, Davi de Oliveira Almeida, Victor Augustus Marin, Marize Pereira Miagostovich
• Format: Article
• Language: English
• Published: Fundação Oswaldo Cruz (FIOCRUZ) 2014-08-01
• Series: Vigilância Sanitária em Debate: Sociedade, Ciência & Tecnologia
• Subjects: Norovirus; Filtration; Lettuce; PP7 Bacteriophage; Internal Control Process
• Online Access: https://hm-visaemdebate.incqs.fiocruz.br/index.php/visaemdebate/article/view/195

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.