Detection and quantification of noroviruses in shellfish.

Noroviruses (NoVs) are the most common viral agents of acute gastroenteritis in humans, and high concentrations of NoVs are discharged into the environment. As these viruses are very resistant to inactivation, the sanitary consequences are contamination of food, including molluscan shellfish. There are four major problems with NoV detection in shellfish samples: low levels of virus contamination, the difficulty of efficient virus extraction, the presence of interfering substances that inhibit molecular detection, and NoV genetic variability.

Norwalk virus-specific binding to oyster digestive tissues.

The primary pathogens related to shellfish-borne gastroenteritis outbreaks are noroviruses. These viruses show persistence in oysters, which suggests an active mechanism of virus concentration. We investigated whether Norwalk virus or viruslike particles bind specifically to oyster tissues after bioaccumulation or addition to tissue sections.

Human and animal enteric caliciviruses in oysters from different coastal regions of the United States.

Food-borne diseases are a major cause of morbidity and hospitalization worldwide. Enteric caliciviruses are capable of persisting in the environment and in the tissues of shellfish. Human noroviruses (HuNoVs) have been implicated in outbreaks linked to shellfish consumption.

Use of rotavirus virus-like particles as surrogates to evaluate virus persistence in shellfish.

Rotavirus virus-like particles (VLPs) and MS2 bacteriophages were bioaccumulated in bivalve mollusks to evaluate viral persistence in shellfish during depuration and relaying under natural conditions. Using this nonpathogenic surrogate virus, we were able to demonstrate that about 1 log10 of VLPs was depurated after 1 week in warm seawater (22 degrees C). Phage MS2 was depurated more rapidly (about 2 log10 in 1 week) than were VLPs, as determined using a single-compartment model and linear regression analysis.

Rotavirus VLP2/6: a new tool for tracking rotavirus in the marine environment.

The potential of rotavirus 2/6-virus-like-particles (VLP2/6) for use as tracers in the marine environment was investigated. The stability of bovine rotavirus (strain RF) and VLP2/6 in natural seawater at 25 degrees C for six days was studied. ELISA and western blot methods were used to quantify the particles.

Rotavirus virus-like particles as surrogates in environmental persistence and inactivation studies.

Virus-like particles (VLPs) with the full-length VP2 and VP6 rotavirus capsid proteins, produced in the baculovirus expression system, have been evaluated as surrogates of human rotavirus in different environmental scenarios. Green fluorescent protein-labeled VLPs (GFP-VLPs) and particles enclosing a heterologous RNA (pseudoviruses), whose stability may be monitored by flow cytometry and antigen capture reverse transcription-PCR, respectively, were used. After 1 month in seawater at 20 degrees C, no significant differences were observed between the behaviors of GFP-VLPs and of infectious rotavirus, whereas pseudovirus particles showed a higher decay rate.

A semiquantitative approach to estimate Norwalk-like virus contamination of oysters implicated in an outbreak.

Gastroenteritis outbreaks linked to shellfish consumption are numerous and Norwalk-like viruses (NLVs) are frequently the responsible causative agents. However, molecular data linking shellfish and clinical samples are still rare despite the availability of diagnostic methods. In a recent outbreak we found the same NLV sequence in stool and shellfish samples (100% identity over 313 bp in the capsid region), supporting the epidemiological data implicating the shellfish as the source of infection.