Comparison of QIAGEN automated nucleic acid extraction methods for CMV quantitative PCR testing.
We examined the effect of nucleic acid extraction methods on the analytic characteristics of a quantitative polymerase chain reaction (PCR) assay for cytomegalovirus (CMV). Human serum samples were extracted with 2 automated instruments (BioRobot EZ1 and QIAsymphony SP, QIAGEN, Valencia, CA) and CMV PCR results compared with those of pp65 antigenemia testing. Both extraction methods yielded results that were comparably linear and precise, whereas the QIAsymphony SP had a slightly lower limit of detection (1.92 log(10) copies/mL vs 2.26 log(10) copies/mL). In both cases, PCR was more sensitive than CMV antigen detection, detecting CMV viremia in 12% (EZ1) and 21% (QIAsymphony) of antigen-negative specimens. This study demonstrates the feasibility of using 2 different extraction techniques to yield results within 0.5 log(10) copies/mL of the mean value, a level that would allow for clinical comparison between different laboratory assays.
Dept of Laboratory Medicine, University of California San Francisco, 94143, USA.
Steve
S
Miller
Henrietta
H
Seet
Yasmeen
Y
Khan
Carolyn
C
Wright
Rohan
R
Nadarajah
D003587:Cytomegalovirus / Q000235:genetics*; D006801:Humans; D057230:Limit of Detection; D009696:Nucleic Acids / Q000032:analysis*; D016133:Polymerase Chain Reaction / Q000295:instrumentation / Q000379:methods*; D012367:RNA, Viral / Q000032:analysis*
D009696:Nucleic Acids; D012367:RNA, Viral