Performance Evaluation of the Real-Q Cytomegalovirus (CMV) Quantification Kit Using Two Real-Time PCR Systems for Quantifying CMV DNA in Whole Blood.
10.3343/alm.2016.36.6.603
- Author:
Jong Eun PARK
1
;
Ji Youn KIM
;
Sun Ae YUN
;
Myoung Keun LEE
;
Hee Jae HUH
;
Jong Won KIM
;
Chang Seok KI
Author Information
1. Department of Laboratory Medicine and Genetics, Samsung Medical Center, Sungkyunkwan University School of Medicine, Seoul, Korea. pmhhj77@gmail.com, changski@skku.edu
- Publication Type:Brief Communication ; Evaluation Studies
- Keywords:
Cytomegalovirus;
Real-time PCR;
Quantification;
Whole blood;
WHO International Standard;
Validation
- MeSH:
Cytomegalovirus/*genetics/isolation & purification;
Cytomegalovirus Infections/diagnosis/virology;
DNA, Viral/*blood/metabolism;
Humans;
Limit of Detection;
Reagent Kits, Diagnostic;
*Real-Time Polymerase Chain Reaction
- From:Annals of Laboratory Medicine
2016;36(6):603-606
- CountryRepublic of Korea
- Language:English
-
Abstract:
Standardized cytomegalovirus (CMV) DNA quantification is important for managing CMV disease. We evaluated the performance of the Real-Q CMV Quantification Kit (Real-Q assay; BioSewoom, Korea) using whole blood (WB), with nucleic acid extraction using MagNA Pure 96 (Roche Diagnostics, Germany). Real-time PCR was performed on two platforms: the 7500 Fast real-time PCR (7500 Fast; Applied Biosystems, USA) and CFX96 real-time PCR detection (CFX96; Bio-Rad, USA) systems. The WHO international standard, diluted with CMV-negative WB, was used to validate the analytical performance. We used 90 WB clinical samples for comparison with the artus CMV RG PCR kit (artus assay; Qiagen, Germany). Limits of detections (LODs) in 7500 Fast and CFX96 were 367 and 479 IU/mL, respectively. The assay was linear from the LOD to 10(6) IU/mL (R2 ≥0.9886). The conversion factors from copies to IU in 7500 Fast and CFX96 were 0.95 and 1.06, respectively. Compared with the artus assay, for values <1,000 copies/mL, 100% of the samples had a variation <0.7 log10 copies/mL; >1,000 copies/mL, 73.3% and 80.6% of samples in 7500 Fast and CFX96, respectively, had <0.5 log10 copies/mL. The Real-Q assay is useful for quantifying CMV in WB with the two real-time PCR platforms.
