Real-Time PCR Assays for the Quantification of HCV RNA: Concordance, Discrepancies and Implications for Response Guided Therapy

Straßl, Robert; Rutter, Karoline; Stättermayer, Albert Friedrich; Beinhardt, Sandra; Kammer, Michael; Hofer, Harald; Ferenci, Péter; Popow‐Kraupp, Theresia

doi:10.1371/journal.pone.0135963

Cited by 18 publications

(10 citation statements)

References 32 publications

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“…Depending on their LLOQ, other assays use lower levels [52–55]. Thus, the threshold and the number of patients achieving virological response may differ depending on the assay and influence clinical management [56–62]. For instance, patients receiving first generation DAA (Telaprevir) combined with PegIFN and RBV and with HCV RNA undetectable at week 4 were used to identify patients eligible for shortened treatment (24 vs 48 weeks) [63].…”

Section: Importance Of Viral Load and Genotyping In The Outcome Of Inmentioning

confidence: 99%

Hepatitis C virus: life cycle in cells, infection and host response, and analysis of molecular markers influencing the outcome of infection and response to therapy

Dustin

Bartolini

Capobianchi

et al. 2016

Clinical Microbiology and Infection

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Hepatitis C virus (HCV) is a major global health burden accounting for around 170 million chronic infections worldwide. Since its discovery, which dates back to about 30 years ago, many details of the viral genome organization and the astonishing genetic diversity have been unveiled but, owing to the difficulty of culturing HCV in vitro and obtaining fully susceptible yet immunocompetent in vivo models, we are still a long way from the full comprehension of viral life cycle, host cell pathways facilitating or counteracting infection, pathogenetic mechanisms in vivo, and host defenses. Here, we illustrate the viral life cycle into cells, describe the interplay between immune and genetic host factors shaping the course of infection, and provide details of the molecular approaches currently used to genotype, monitor replication in vivo, and studying the emergence of drug-resistant viral variants.

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Section: Importance Of Viral Load and Genotyping In The Outcome Of Inmentioning

confidence: 99%

Hepatitis C virus: life cycle in cells, infection and host response, and analysis of molecular markers influencing the outcome of infection and response to therapy

Dustin

Bartolini

Capobianchi

et al. 2016

Clinical Microbiology and Infection

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“…The performances of some of these existing HCV quantitative assays have been compared side by side in various published studies ( 13 , 23 – 26 ). However, the Aptima assay has not yet been evaluated in comparison to other assays.…”

Section: Introductionmentioning

confidence: 99%

Performance of the New Aptima HCV Quant Dx Assay in Comparison to the Cobas TaqMan HCV2 Test for Use with the High Pure System in Detection and Quantification of Hepatitis C Virus RNA in Plasma or Serum

Schalasta¹,

Speicher²,

Börner³

et al. 2016

J Clin Microbiol

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Quantitating the level of hepatitis C virus (HCV) RNA is the standard of care for monitoring HCV-infected patients during treatment. The performances of commercially available assays differ for precision, limit of detection, and limit of quantitation (LOQ). Here, we compare the performance of the Hologic Aptima HCV Quant Dx assay (Aptima) to that of the Roche Cobas TaqMan HCV test, version 2.0, using the High Pure system (HPS/CTM), considered a reference assay since it has been used in trials defining clinical decision points in patient care. The assays' performance characteristics were assessed using HCV RNA reference panels and plasma/serum from chronically HCV-infected patients. The agreement between the assays for the 3 reference panels was good, with a difference in quantitation values of <0.5 log. High concordance was demonstrated between the assays for 245 clinical samples (kappa = 0.80; 95% confidence interval [CI], 0.720 to 0.881); however, Aptima detected and/or quantitated 20 samples that HPS/CTM did not detect, while Aptima did not detect 1 sample that was quantitated by HPS/CTM. For the 165 samples quantitated by both assays, the values were highly correlated (R = 0.98; P < 0.0001). The linearity of quantitation from concentrations of 1.4 to 6 log was excellent for both assays for all HCV genotypes (GT) tested (GT 1a, 1b, 2b, and 3a) (R2 > 0.99). The assays had similar levels of total and intra-assay variability across all genotypes at concentrations from 1,000 to 25 IU/ml. Aptima had a greater analytical sensitivity, quantitating more than 50% of replicates at 25-IU/ml target. Aptima showed performance characteristics comparable to those of HPS/CTM and increased sensitivity, making it suitable for use as a clinical diagnostic tool on the fully automated Panther platform.

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“…94,95 The problem of discordant results between different platforms becomes worse in the case of low viral loads. 96 Nested qRT-PCR can increase detection sensitivity; however, it is more sensitive to contamination and has higher false-positive rates. 97 Droplet digital PCR was recently suggested as a minimal sample preparation substitute for qRT-PCR; but it remains highly equipment/personnel intensive.…”

Section: Quantitative Reverse Transcription-pcr (Qrt-pcr)mentioning

confidence: 99%

Future microfluidic and nanofluidic modular platforms for nucleic acid liquid biopsy in precision medicine

et al. 2016

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Nucleic acid biomarkers have enormous potential in non-invasive diagnostics and disease management. In medical research and in the near future in the clinics, there is a great demand for accurate miRNA, mRNA, and ctDNA identification and profiling. They may lead to screening of early stage cancer that is not detectable by tissue biopsy or imaging. Moreover, because their cost is low and they are non-invasive, they can become a regular screening test during annual checkups or allow a dynamic treatment program that adjusts its drug and dosage frequently. We briefly review a few existing viral and endogenous RNA assays that have been approved by the Federal Drug Administration. These tests are based on the main nucleic acid detection technologies, namely, quantitative reverse transcription polymerase chain reaction (PCR), microarrays, and next-generation sequencing. Several of the challenges that these three technologies still face regarding the quantitative measurement of a panel of nucleic acids are outlined. Finally, we review a cluster of microfluidic technologies from our group with potential for point-of-care nucleic acid quantification without nucleic acid amplification, designed to overcome specific limitations of current technologies. We suggest that integration of these technologies in a modular design can offer a low-cost, robust, and yet sensitive/selective platform for a variety of precision medicine applications.

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Real-Time PCR Assays for the Quantification of HCV RNA: Concordance, Discrepancies and Implications for Response Guided Therapy

Cited by 18 publications

References 32 publications

Hepatitis C virus: life cycle in cells, infection and host response, and analysis of molecular markers influencing the outcome of infection and response to therapy

Hepatitis C virus: life cycle in cells, infection and host response, and analysis of molecular markers influencing the outcome of infection and response to therapy

Performance of the New Aptima HCV Quant Dx Assay in Comparison to the Cobas TaqMan HCV2 Test for Use with the High Pure System in Detection and Quantification of Hepatitis C Virus RNA in Plasma or Serum

Future microfluidic and nanofluidic modular platforms for nucleic acid liquid biopsy in precision medicine

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