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  1. Sensitive detection of group A rotaviruses by immunomagnetic separation and reverse transcription-polymerase chain reaction.

    Article - En anglais

    An immunomagnetic separation (IMS) method was developed for concentrating rotaviruses from environmental samples, as well as a reverse transcription-polymerase chain reaction (RT-PCR) for the sensitive and specific detection of group A rotaviruses.

    Magnetic beads were coated with monoclonal antibodies directed against the group-specific, inner capsid protein (VP6) and subsequently used to capture and purify the virus with the help of a magnet.

    The genome was made available for RT by heat-disrupting the viral particles.

    A single 40-cycle PCR was as sensitive as a nested PCR, both detecting 0.005 PFU of the Wa strain, corresponding to approximately 5 particles as indicated by EM.

    The nested PCR was positive for all the group A strains tested, but negative for group C rotaviruses and other RNA viruses.

    The IMS-RT-PCR method functioned satisfactorily with virus seeded out in fresh water samples ; with sea water, the IMS removed most, but not all, inhibiting activity.

    Mots-clés Pascal : Rotavirus, Reoviridae, Virus, Détection, Méthode, Transcription inverse, Réaction chaîne polymérase, Evaluation performance, Anticorps monoclonal, Méthode immunomagnétique

    Mots-clés Pascal anglais : Rotavirus, Reoviridae, Virus, Detection, Method, Reverse transcription, Polymerase chain reaction, Performance evaluation, Monoclonal antibody, Immunomagnetic method

    Logo du centre Notice produite par :
    Inist-CNRS - Institut de l'Information Scientifique et Technique

    Cote : 96-0071908

    Code Inist : 002A05C09. Création : 199608.