Immunomagnetic separation of a Norwalk-like virus (genogroup I) in artificially contaminated environmental water samples

Rabbit polyclonal antibodies were raised against a recombinant capsid prote in from a genogroup I Norwalk-like virus (NLV). Magnetic beads coated with these antibodies were used in immunomagnetic separation (IMS) of the NLV. A fter capture of the NLV and washing of the beads, viral RNA was heat releas ed and detected by RT-PCR. This IMS procedure was shown to have high sensit ivity for detection of homologous NLV, while capture of a genogroup II NLV was less efficient. Antigen capture was not influenced by the content of hu mic acids in the samples. The combination of IMS and heat release was found to be more efficient than organic extraction of RNA from water contaminate d with humic acids. The efficacy and simplicity of IMS/heat release render this combination a feasible tool for the preparation of NLV RNA from enviro nmental samples, although the antigenic diversity of NLV may be a complicat ing factor. (C) 2000 Elsevier Science B.V. All rights reserved.