Automated Purification of Viral Nucleic Acid from Liquid Samples for Qualitative...

Abstract

Rachel Mann¹, Rob Christensen², Keisha Church³ and Trista Schagat^1
1Promega Corporation, Madison, WI USA
²Wyoming Public Health Lab, Cheyenne, WY USA
³Medical University of South Carolina, Charleston, SC USA

Viruses vary in coat structure, nucleic acid content and body fluids in which they can be found. This can necessitate different extraction methods for different viruses. Here we describe a simple protocol for automated isolation of viral DNA and RNA in liquid samples using the prefilled cartridges of the Maxwell^® LEV Total RNA Purification Kit. Viral nucleic acid was successfully isolated and detected from both RNA (norovirus, influenza, HCV, HIV) and DNA (CMV, HSV) viruses in a range of fluids (plasma, medium from nasal swabs, fecal supernatant). Detection was based on amplification and confirmed by a second extraction method. The protocol presented is recommended for qualitative detection only, as the efficiency of recovery remains to be determined.