title: Automated Nucleic Acid Extraction Systems for Detecting Cytomegalovirus and Epstein-Barr Virus Using Real-Time PCR: A Comparison Study Between the QIAsymphony RGQ and QIAcube Systems.
abstract: BACKGROUND
  Cytomegalovirus (CMV) and Epstein-Barr virus (EBV) are increasingly important in immunocompromised patients. Nucleic acid extraction methods could affect the results of viral nucleic acid amplification tests. We compared two automated nucleic acid extraction systems for detecting CMV and EBV using real-time PCR assays.
  
  
  METHODS
  One hundred and fifty-three whole blood (WB) samples were tested for CMV detection, and 117 WB samples were tested for EBV detection. Viral nucleic acid was extracted in parallel by using QIAsymphony RGQ and QIAcube (Qiagen GmbH, Germany), and real-time PCR assays for CMV and EBV were performed with a Rotor-Gene Q real-time PCR cycler (Qiagen). Detection rates for CMV and EBV were compared, and agreements between the two systems were analyzed.
  
  
  RESULTS
  The detection rate of CMV and EBV differed significantly between the QIAsymphony RGQ and QIAcube systems (CMV, 59.5% [91/153] vs 43.8% [67/153], P=0.0005; EBV, 59.0% [69/117] vs 42.7% [50/117], P=0.0008). The two systems showed moderate agreement for CMV and EBV detection (kappa=0.43 and 0.52, respectively). QIAsymphony RGQ showed a negligible correlation with QIAcube for quantitative EBV detection. QIAcube exhibited EBV PCR inhibition in 23.9% (28/117) of samples.
  
  
  CONCLUSIONS
  Automated nucleic acid extraction systems have different performances and significantly affect the detection of viral pathogens. The QIAsymphony RGQ system appears to be superior to the QIAcube system for detecting CMV and EBV. A suitable sample preparation system should be considered for optimized nucleic acid amplification in clinical laboratories.
authors.affiliation: Department of Laboratory Medicine, Konkuk University School of Medicine, Seoul, Korea.
authors.forename: Hanah
authors.initials: H
authors.lastname: Kim
authors.affiliation: Department of Laboratory Medicine, Konkuk University School of Medicine, Seoul, Korea. dearmina@hanmail.net.
authors.forename: Mina
authors.initials: M
authors.lastname: Hur
authors.affiliation: Department of Laboratory Medicine, Konkuk University School of Medicine, Seoul, Korea.
authors.forename: Ji Young
authors.initials: JY
authors.lastname: Kim
authors.affiliation: Department of Laboratory Medicine, Konkuk University School of Medicine, Seoul, Korea.
authors.forename: Hee Won
authors.initials: HW
authors.lastname: Moon
authors.affiliation: Department of Laboratory Medicine, Konkuk University School of Medicine, Seoul, Korea.
authors.forename: Yeo Min
authors.initials: YM
authors.lastname: Yun
authors.affiliation: Department of Laboratory Medicine, Hallym University College of Medicine, Seoul, Korea.
authors.forename: Hyun Chan
authors.initials: HC
authors.lastname: Cho
mesh_terms: D001331:Automation; D003587:Cytomegalovirus / Q000235:genetics* / Q000302:isolation & purification; D003586:Cytomegalovirus Infections / Q000175:diagnosis / Q000821:virology*; D004279:DNA, Viral / Q000097:blood* / Q000302:isolation & purification / Q000378:metabolism; D004854:Herpesvirus 4, Human / Q000235:genetics* / Q000302:isolation & purification; D006801:Humans; D011933:Reagent Kits, Diagnostic; D060888:Real-Time Polymerase Chain Reaction
chemical_list: D004279:DNA, Viral; D011933:Reagent Kits, Diagnostic
keywords: Cytomegalovirus; Epstein-Barr virus; Extraction; Nucleic acid; Performance; QIAcube; QIAsymphony RGQ