Transcriptional enhancement of RT-PCR for rapid and sensitive detection of Noroviruses

Previously reported nucleic acid sequence-based amplification (NASBA) primers specific for the GII Noroviruses were adapted for reverse transcriptase-polymerase chain reaction (RT-PCR), and detection sensitivity was then enhanced by a subsequent in vitro transcription of the RT-PCR amplicons. The NASBA-derived primers performed comparably to other broadly reactive GII Norovirus primers with respect to detection limits (i.e. I RT-PCR amplifiable unit (RT-PCRU) per reaction). Detection limits improved by approximately I log(10) to 0.3 RT-PCRU per reaction when transcriptional enhancement and electrochemiluminescence (ECL) hybridization followed RT-PCR. The method shows promise for improved detection sensitivity in instances where very low levels of virus contamination might be anticipated. (C) 2003 Federation of European Microbiological Societies. Published by Elsevier B.V. All rights reserved.