Comparison of seven RNA extraction methods on stool and shellfish samples prior to hepatitis A virus amplification

When choosing an extraction method, two parameters have to be considered: r ecovery of the viral material and elimination or inactivation of inhibitory substances. Seven techniques for extracting hepatitis A virus (HAV) from s tool and shellfish samples were compared, in order to identify the protocol most suited to both types of sample and with the best extraction yield. Th e protocols tested were either techniques for the recovery and purification of total RNA, such as RNAzol, PEG-CETAB, GTC-silica and Chelex, or techniq ues for isolating specifically HAV using a nucleotide probe or a monoclonal antibody. For stool samples, RNAzol, PEG-CETAB, and magnetic beads with an tibody allowed detection of the virus in 11/12 and 12/12 of samples. For sh ellfish samples, three protocols allowed RNA to be extracted in 90% of case s, RNAzol, PEG-CETAB, and GTC-silica. Their rapidity and low cost make RNAz ol and GTC-silica the most suitable for routine diagnostic testing. (C) 199 9 Elsevier Science B.V. All rights reserved.