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Recovery efficiences on nucleic acid extraction kits as measured by quantitative LightCycler PCR
1Micropathology Ltd, University of Warwick, Coventry, UK. steve.read@micropathology.com
Aims:
To compare the efficiency of five nucleic acid extraction kits for processing clinical material for the diagnosis of infection.
Methods:
Five nucleic acid extraction kits for processing clinical material for the diagnosis of infection were compared for their relative efficiencies in purifying and recovering either viral DNA or RNA from serum samples. Quantitative polymerase chain reaction (PCR) assays for hepatitis B virus (HBV) DNA and hepatitis C virus (HCV) RNA were performed on the Light-Cycler instrument to determine the relative concentrations of the viral nucleic acids recovered by the various protocols.
Results:
Large differences between kits in recovery efficiencies were observed for HBV DNA, with those protocols using enzyme digestion in addition to chaotropic solutions performing better than those using chemical disruption alone.
Conclusions:
There were large differences between the kits and it appeared that those extraction kits containing a proteolytic enzyme and carrier nucleic acid (and that are supplied RNase free) have the widest potential application in the routine microbiology laboratory.