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

• Published in: Annals of laboratory medicine Vol. 37; no. 2; pp. 129 - 136
• Main Authors: Kim, Hanah, Hur, Mina, Kim, Ji Young, Moon, Hee Won, Yun, Yeo Min, Cho, Hyun Chan
• Format: Journal Article
• Language: English
• Published: Korea (South) The Korean Society for Laboratory Medicine 01-03-2017; 대한진단검사의학회
• Subjects: Automation; Cytomegalovirus - genetics; Cytomegalovirus - isolation & purification; Cytomegalovirus Infections - diagnosis; Cytomegalovirus Infections - virology; DNA, Viral - blood; DNA, Viral - isolation & purification; DNA, Viral - metabolism; Herpesvirus 4, Human - genetics; Herpesvirus 4, Human - isolation & purification; Humans; Original; Reagent Kits, Diagnostic; Real-Time Polymerase Chain Reaction; 병리학; Cytomegalovirus; QIAcube; QIAsymphony RGQ; Epstein-Barr virus; Extraction; Performance; Nucleic acid
• ISSN: 2234-3806; 2234-3814
• DOI: 10.3343/alm.2017.37.2.129

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.
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.
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.
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.