Immunomagnetic separation of a Norwalk-like virus (genogroup I) in artificially contaminated environmental water samples.
Rabbit polyclonal antibodies were raised against a recombinant capsid protein from a genogroup I Norwalk-like virus (NLV). Magnetic beads coated with these antibodies were used in immunomagnetic separation (IMS) of the NLV. After capture of the NLV and washing of the beads, viral RNA was heat released and detected by RT-PCR. This IMS procedure was shown to have high sensitivity for detection of homologous NLV, while capture of a genogroup II NLV was less efficient. Antigen capture was not influenced by the content of humic acids in the samples. The combination of IMS and heat release was found to be more efficient than organic extraction of RNA from water contaminated with humic acids. The efficacy and simplicity of IMS/heat release render this combination a feasible tool for the preparation of NLV RNA from environmental samples, although the antigenic diversity of NLV may be a complicating factor.
Myrmel
M
M
Department of Pharmacologv, Microbiology and Food Hygiene, The Norwegian School of Veterinary Science, Oslo. mette.myrmel@veths.no
Rimstad
E
E
Wasteson
Y
Y
D000818:Animals; D000914:Antibodies, Viral / Q000097:blood; D002213:Capsid / Q000276:immunology*; D002614:Chelating Agents; D006358:Hot Temperature; D006812:Humic Substances; D018189:Immunomagnetic Separation / Q000379:methods*; D009663:Norwalk virus / Q000235:genetics / Q000276:immunology / Q000302:isolation & purification*; D012367:RNA, Viral / Q000032:analysis; D011817:Rabbits; D011994:Recombinant Proteins / Q000276:immunology; D020133:Reverse Transcriptase Polymerase Chain Reaction; D012680:Sensitivity and Specificity; D014871:Water Microbiology
D000914:Antibodies, Viral; D002614:Chelating Agents; D006812:Humic Substances; D012367:RNA, Viral; D011994:Recombinant Proteins