Comparison of magnetic bead and rapid swab RNA extraction methods for detecting rabbit hemorrhagic disease virus 2 in rabbit liver samples

Abstract

We compared a bead RNA extraction method with a one-tube method that required only a heat block and ice. RNA was first extracted from liver samples from nine rabbits dying from rabbit hemorrhagic disease virus 2 (RHDV2) using magnetic beads, and RT-PCR was used to detect RHDV2 sequence. Following freezing, RNA was extracted a second time using the SwiftX™ Swabs Viral RNA Extraction Reagent. RHDV2 was detected in all nine samples. Cycle threshold values were higher in the RT-PCR following SwiftX extraction (mean: 3.79), indicating that the second extraction method resulted in approximately a 1 log10 reduction in sensitivity. A second freeze–thaw for the samples and less tissue extracted using SwiftX may have contributed additionally to the loss in sensitivity.

Keywords

• field studies
• rabbit hemorrhagic disease virus 2
• rapid nucleic extraction methods
• RHDV2
• RNA extraction
• RNA virus detection