Comparison of two nucleic acid extraction and testing systems for HCMV-DNA detection and quantitation on whole blood specimens from transplant patients.
Quantitative detection of human cytomegalovirus (HCMV) DNA on whole blood is currently the primary choice for virological monitoring in transplant patients and for determining the appropriate antiviral strategy, however specific issues of variability remain in terms of extraction methods, amplification efficiency, and variability. This study compared the performance characteristics of two nucleic acid extraction and testing systems for HCMV-DNA quantitation, the artus® CMV QS-RGQ kit, associated with a fully automated DNA extraction and assay set up by Qiagen (system 1) and the Q-CMV Real Time Complete kit by Nanogen, associated with a semiautomated nucleic acid extraction system by Biomérieux (system 2) in 189 specimens from transplant patients and 10 from 2012 HCMV Quality Control for Molecular Diagnostics (QCMD). The two systems exhibited a 80.4% concordance. Differences between the two systems were within ±1 log10 copies/ml of the averaged log10 results for 88.9% of the tested specimens. For all qualitatively discordant specimens, mean viral load was ≤3 log10 copies/ml. Considering viral load measurement, system 1 gave earlier positives that system 2, with a 14.8% of specimens resulted positive at low viral loads with system 1 and negative with system 2. In QCMD specimens, difference was below 0.7 log10 copies/ml for both the systems. In conclusion, the two systems provided reliable and comparable results. Some specific performance characteristic and automation could be taken into account in terms of less hands of time, fewer errors and reliability.
Microbiology and Virology Unit, Turin, Italy. Electronic address: cristina.costa@unito.it.
Cristina
C
Costa
Samantha
S
Mantovani
Cinzia
C
Balloco
Francesca
F
Sidoti
Fabrizio
F
Fop
Rossana
R
Cavallo
D057205:Automation, Laboratory / Q000379:methods; D001769:Blood / Q000821:virology*; D003587:Cytomegalovirus / Q000235:genetics / Q000302:isolation & purification*; D003586:Cytomegalovirus Infections / Q000175:diagnosis* / Q000821:virology; D004279:DNA, Viral / Q000235:genetics / Q000302:isolation & purification*; D006801:Humans; D025202:Molecular Diagnostic Techniques / Q000379:methods*; D060888:Real-Time Polymerase Chain Reaction / Q000379:methods*; D013048:Specimen Handling / Q000379:methods*; D014180:Transplantation; D019562:Viral Load / Q000379:methods
D004279:DNA, Viral
Efficiency; Extraction; Human cytomegalovirus; Nucleic acid; Quantitative PCR; Variability