Performance Characteristics of a Quantitative, Homogeneous TaqMan RT-PCR Test for HCV RNA Joerg Kleiber, Thomas Walter, Gerd Haberhausen, Sue Tsang,

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Presentation transcript:

Performance Characteristics of a Quantitative, Homogeneous TaqMan RT-PCR Test for HCV RNA Joerg Kleiber, Thomas Walter, Gerd Haberhausen, Sue Tsang, Reiner Babiel, Maurice Rosenstraus The Journal of Molecular Diagnostics Volume 2, Issue 3, Pages 158-166 (August 2000) DOI: 10.1016/S1525-1578(10)60632-0 Copyright © 2000 American Society for Investigative Pathology and Association for Molecular Pathology Terms and Conditions

Figure 1 Efficiency of amplification of different HCV genotypes. HCV transcripts representing genotypes 1a, 1b, 2a, 2b, 3a, 4, and 5 were diluted to 100,000 copies/ml and tested with the TaqMan HCV quantitative Test. The CT for four replicates of each genotype were averaged to determine the mean CT and SD. The Journal of Molecular Diagnostics 2000 2, 158-166DOI: (10.1016/S1525-1578(10)60632-0) Copyright © 2000 American Society for Investigative Pathology and Association for Molecular Pathology Terms and Conditions

Figure 2 Linear range for the TaqMan HCV test. Two independent dilution series were prepared by diluting an external HCV RNA quantitation standard in HCV-negative plasma. Each dilution in each series was tested in duplicate. At each input, RNA concentration, the mean of the log10 calculated RNA concentrations (circles and squares), and the SD were determined. Linear regression analysis was performed on the mean log10 values for each dilution series (solid and dashed lines and associated equations). The Journal of Molecular Diagnostics 2000 2, 158-166DOI: (10.1016/S1525-1578(10)60632-0) Copyright © 2000 American Society for Investigative Pathology and Association for Molecular Pathology Terms and Conditions

Figure 3 Quantitation of HCV RNA (left axis) and serum ALT (right axis) levels in a responder-relapser (A), responder (B), and nonresponder (C) to interferon therapy. Baseline HCV RNA and ALT levels (first data point) were measured 12 (A), 4 (B), and 65 (C) weeks before the start of interferon therapy (day 0,vertical dashed line). RNA and ALT levels were periodically monitored starting at 1 week after initiating therapy (second data point). The posttreatment RNA levels (last data point) were measured at 25 (A), 4 (B) or 38 (C) weeks after completion of interferon therapy at week 48 (vertical dashed line). The dotted-dashed lineindicates both the limit of detection for the HCV RNA test and the upper limit of the normal ALT range. Samples that tested negative for HCV RNA were assigned an arbitrary value (10 IU/ml) below the limit of detection. The Journal of Molecular Diagnostics 2000 2, 158-166DOI: (10.1016/S1525-1578(10)60632-0) Copyright © 2000 American Society for Investigative Pathology and Association for Molecular Pathology Terms and Conditions