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Journal of clinical microbiology v.55 no.2, 2017년, pp.504 - 509   SCI SCIE
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Assessment of a Novel Automatic Real-Time PCR Assay on the Cobas 4800 Analyzer as a Screening Platform for Hepatitis C Virus Genotyping in Clinical Practice: Comparison with Massive Sequencing

Nieto-Aponte, Leonardo (Liver Pathology Unit, Departments of Biochemistry and Microbiology, Hospital Universitari Vall d'Hebron (HUVH), Universitat Autònoma de Barcelona (UAB), Barcelona, Spain ) ; Quer, Josep (Centro de Investigaciòón Biomòóédica en Red de Enfermedades Hepòóéáticas y Digestivas (CIBERehd), Instituto de Salud Carlos III, Madrid, Spain ) ; Ruiz-Ripa, Alicia (Liver Pathology Unit, Departments of Biochemistry and Microbiology, Hospital Universitari Vall d'Hebron (HUVH), Universitat Autònoma de Barcelona (UAB), Barcelona, Spain ) ; Tabernero, David (Liver Pathology Unit, Departments of Biochemistry and Microbiology, Hospital Universitari Vall d'Hebron (HUVH), Universitat Autònoma de Barcelona (UAB), Barcelona, Spain ) ; Gonzalez, Carolina (Liver Pathology Unit, Departments of Biochemistry and Microbiology, Hospital Universitari Vall d'Hebron (HUVH), Universitat Autònoma de Barcelona (UAB), Barcelona, Spain ) ; Gregori, Josep (Centro de Investigaciòón Biomòóédica en R ) ; Vila, Marta ; Asensio, Miriam ; Garcia-Cehic, Damir ; Ruiz, Gerardo ; Chen, Qian ; Ordeig, Laura ; Llorens, Meritxell ; Saez, Montserrat ; Esteban, Juan I. ; Esteban, Rafael ; Buti, Maria ; Pumarola, Tomas ; Rodriguez-Frias, Francisco ;
  • 초록  

    The unequivocal identification of hepatitis C virus (HCV) subtypes 1a/1b and genotypes 2 to 6 is required for optimizing the effectiveness of interferon-free, direct-acting antiviral therapies. We compared the performance of a new real-time HCV genotyping assay used on the Cobas 4800 system (C4800) with that of high-resolution HCV subtyping (HRCS). In total, 502 samples were used, including 184 samples from chronic HCV patients (from routine laboratory activity during April 2016), 5 stored samples with double HCV genotype infections for testing the limitations of the method, and 313 samples from a screening protocol implemented in our hospital (from May to August 2016) based on the new method to further determine its genotyping accuracy. A total of 282 samples, including 171 from April 2016 (the 13 remaining had too low of a viral load for HRCS), 5 selected with double infections, and 106 from screening, were analyzed by both methods, and 220 were analyzed only by the C4800. The C4800 correctly subtyped 125 of 126 1a/1b samples, and the 1 remaining sample was reported as genotype 1. The C4800 correctly genotyped 38 of 45 non-1a/1b samples (classified by HRCS), and it reported the remaining 7 samples as indeterminate. One hundred two of 106 non-1a/1b genotype samples that were identified using the C4800 for screening were confirmed by HRCS. In the 4 remaining samples, 3 were correctly reported as genotype 1 (without defining the subtype) and 1 was reported as indeterminate. None of the samples were misgenotyped. Four of 7 samples with double HCV infections were correctly genotyped by the C4800. Excluding the 5 selected double-infected samples, the C4800 showed 95.7% concordant results for genotyping HCVs 2 to 6 and 1a/1b subtyping, and 99.2% concordance for subtyping 1a/1b single infections in clinical samples. To improve laboratory workflow, we propose using the C4800 as a first-line test for HCV genotyping and 1a/1b classification, followed by transferring non-1a/1b samples to a center where HRCS is available, if further characterization is needed.


  • 주제어

    diagnostics .   genotype identification .   hepatitis C virus .   molecular subtyping.  

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