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
| Autor: | Hee Won Moon, Jiyoung Kim, Mina Hur, Hanah Kim, Hyun Chan Cho, Yeo-Min Yun |
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| Rok vydání: | 2017 |
| Předmět: |
QIAsymphony RGQ
0301 basic medicine Herpesvirus 4 Human Performance Clinical Biochemistry Pcr assay Cytomegalovirus Extraction QIAcube Biology Real-Time Polymerase Chain Reaction medicine.disease_cause Virus Automation 03 medical and health sciences medicine Humans Epstein-Barr virus Biochemistry (medical) General Medicine Epstein–Barr virus Virology 030104 developmental biology Real-time polymerase chain reaction Nucleic acid Cytomegalovirus Infections DNA Viral Comparison study Original Article Extraction methods Reagent Kits Diagnostic Diagnostic Genetics |
| Zdroj: | Annals of Laboratory Medicine |
| ISSN: | 2234-3814 2234-3806 |
| Popis: | 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. |
| Databáze: | OpenAIRE |
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