Please cite this article in press as: Paba, P., et al., Performance evaluation of the Artus hepatitis C virus QS-RGQ assay. J. Virol. Methods (2011), doi:10.1016/j.jviromet.2011.09.024 ARTICLE IN PRESS G Model VIRMET-11668; No. of Pages 4 Journal of Virological Methods xxx (2011) xxx–xxx Contents lists available at SciVerse ScienceDirect Journal of Virological Methods jou rn al h om epage: www.elsevier.com/locate/jviromet Performance evaluation of the Artus hepatitis C virus QS-RGQ assay Pierpaolo Paba, Lavinia Fabeni, Carlo Federico Perno, Marco Ciotti ∗ Laboratory of Molecular Virology, Foundation Polyclinic Tor Vergata, Viale Oxford, 81-00133 Rome, Italy Article history: Received 19 July 2011 Received in revised form 21 September 2011 Accepted 28 September 2011 Available online xxx Keywords: HCV real-time PCR HCV viral load a b s t r a c t Accurate determination of hepatitis C virus RNA level is essential for evaluating the response to antiviral therapy, to determine the duration of treatment, and to predict treatment outcome. Currently, two real- time based polymerase chain reaction assays are used widely to monitor the hepatitis C RNA level: the Abbott RealTime HCV assay and the Cobas Taqman HCV assay. Recently, a third assay has become commercially available: the Artus HCV QS-RGQ assay, which uses the QIAsymphony SP/AS platform for sample preparation and PCR-setup, and the Rotor-Gene Q for amplification and detection. In this study, the performance of the Artus HCV QS-RGQ assay was tested on 105 plasma samples and compared to that of the Cobas Taqman HCV assay. Linear regression analysis showed a good agreement between the two assays. A slightly better sensitivity was observed with the Cobas Taqman assay, while higher hepatitis C viral RNA levels were measured by the Artus HCV QS-RGQ assay in samples positive for hepatitis C genotypes 4. Taken together, the data suggest that the Artus HCV QS-RGQ assay is useful in a diagnostic setting. The combination with the versatile QIAsymphony SP/AS system may represent a major advantage for clinical virological laboratories aiming at optimizing their workflow. © 2011 Elsevier B.V. All rights reserved. 1. Introduction Hepatitis C virus (HCV) is the major cause of chronic hepatitis, cirrhosis of the liver and hepatocellular carcinoma in developed countries. HCV genotyping, measurement of viral load and liver fibrosis at the time of diagnosis are used to determine the dura- tion of antiviral therapy and to predict the response to treatment (EASL Clinical Practice Guidelines: Management of hepatitis C virus infection, 2011). Real-time PCR is the method of choice for mea- suring HCV viral load and it offers a series of advantages over the conventional molecular methods: such as (i) increased analytical sensitivity (10–15 IU/ml), (ii) faster results, (iii) reduced risk of con- tamination, and (iv) wider dynamic range (up to 7–8 log IU/ml). Currently, two real-time PCR methods are used for quantitating HCV-RNA: the Cobas Ampliprep/Cobas TaqMan HCV assay (Roche Molecular System, Pleasanton, CA) and the Abbott RealTime HCV assay (Abbott Diagnostics, Chicago, IL) which uses the m2000 sys- tem, a platform capable of automated RNA extraction and PCR set-up, followed by amplification and detection. Both platforms are completely automated but the two systems do not allow to use the extracted nucleic acid for tests other than HCV or to perform more tests simultaneously on the same extract. Recently, a new real-time PCR based assay is available for quantitating HCV-RNA level in plasma samples: the Artus HCV QS-RGQ kit which is run on ∗ Corresponding author. E-mail address: marco.ciotti@ptvonline.it (M. Ciotti). the Rotor-Gene Q thermal cycler associated to the QIAsymphony SP/AS platform (Qiagen, Hilden, Germany), which is dedicated to nucleic acids extraction and PCR-set up. In this study, the perfor- mance of the Artus HCV QS-RGQ assay was compared to that of the Cobas Taqman HCV assay which is routinely used in our laboratory. Unlike the Roche and Abbott platforms, the Qiagen platform allows simultaneous extraction of DNA and RNA from the same sample, the use of the extracted nucleic acid for different tests, even at dif- ferent times, and to perform simultaneously on the same sample more than one test. 2. Materials and methods 2.1. Sample collection The study was carried out on 105 plasma samples collected from 90 patients with chronic hepatitis C attending the Hepatology Unit of the Polyclinic Tor Vergata, Rome, from August 2010 to January 2011. Blood samples were collected in a BD Vacutainer tube con- taining a separation gel. After centrifugation at 839 × g for 10 min, plasma was divided into two aliquots of 1.1 ml each and stored at -80 ◦ C until analysis. Samples were thawed and tested simultane- ously by the Cobas Ampliprep/Cobas TaqMan HCV and the Artus HCV QS-RGQ assays, respectively. The Cobas TaqMan instrumenta- tion used in this study included a docking station. HCV genotype was available for 103 out of 105 samples tested. The mean age of the patients enrolled in this study was 57.7 years, and the female/male ratio was 0.57 (33 females, and 57 0166-0934/$ – see front matter © 2011 Elsevier B.V. All rights reserved. doi:10.1016/j.jviromet.2011.09.024