1. Absolute Quantitation of Different Genotypes of Hepatitis C Virus RNA in Clinical Samples by a Modified Real-Time PCR Method.
- Author
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Trujillo-Murillo, Karina del Carmen, Pérez-Ibave, Diana Cristina, Ríos-Ibarra, Clara Patricia, Ramirez-Valles, Eda Guadalupe, Rincón-Sánchez, Ana Rosa, and Rivas-Estilla, Ana María
- Subjects
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POLYMERASE chain reaction methodology , *CHRONIC diseases , *GENES , *HEPATITIS C , *HEPATITIS viruses , *RESEARCH methodology , *MICROBIAL genetics , *PROBABILITY theory , *REGRESSION analysis , *DATA analysis software ,RESEARCH evaluation - Abstract
Background: Our aim was to develop a quantitative real-time polymerase chain reaction (qPCR) assay for the quantitation of hepatitis C virus (HCV) RNA samples from patients infected with different HCV genotypes. Methods: A standard curve was generated by amplification of serial dilutions of HCV-plasmid (pFKI389-NS3-3′) harboring genotype 1b HCV-subgenomic replicon. Samples from 15 HCV-infected patients (genotypes 1, 2, and 3) were analyzed to quantify HCV-RNA by qPCR with primers and probes specific for the 5'-UTR viral region. Results: The HCV qPCR assay had a sensitivity of 100 copies/reaction with a dynamic range of detection between 102-20 × 106 HCV copies. The assay was highly reproducible with a low coefficient of variation. We observed that the HCV genotypes included could be identified by our method. Conclusions: Our results showed that this modified qPCR assay provides a valid platform for quantifying HCV-RNA, combining good analytical sensitivity with a wide dynamic range and high reproducibility. [ABSTRACT FROM AUTHOR]
- Published
- 2011
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