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Amplification of NS3 and NS5A in hepatitis C virus by the multiplex nested polymerase chain reaction

  • Authors:
    • Anchalee Sistayanarain
    • Duangkamol Kunthalert
  • View Affiliations / Copyright

    Affiliations: Department of Microbiology and Parasitology, Faculty of Medical Science, Naresuan University, Phitsanulok 65000, Thailand
    Copyright: © Sistayanarain et al. This is an open access article distributed under the terms of Creative Commons Attribution License [CC BY 4.0].
  • Article Number: 54
    |
    Published online on: April 7, 2025
       https://doi.org/10.3892/wasj.2025.342
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Abstract

Resistance‑associated substitutions in patients with hepatitis C virus infection have been shown to decrease the efficacy of direct acting antivirals and lead to treatment failure. Amino acid mutations in the NS3 and NS5A regions have been found to be associated with the treatment failure of NS3 and NS5A inhibitors. A method enabling the detection of these mutations would be useful for identifying patients who are at risk of treatment failure. The standard method used to detect mutations is based on nested polymerase chain reaction and nucleotide and amino acid sequencing. The aim of the present study was to simultaneously amplify NS3 and NS5A of HCV by using the multiplex nested PCR method. A set of primers was designed and optimized specifically for the NS3 and NS5A regions, and the multiplex nested PCR conditions were also optimized. A total of 20 serum samples were processed to evaluate the validity of this method. Both the amplified NS3 and NS5A fragments were detected within 14 of the samples (70%) using optimized multiplex nested PCR. Both specific amplification fragments could be detected in a cDNA dilution as low as 1:8. On the whole, the present study demonstrates that optimized multiplex nested PCR is a simple, rapid and effective method that accomplishes the simultaneous amplification of NS3 and NS5A in HCV.
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Copy and paste a formatted citation
Spandidos Publications style
Sistayanarain A and Kunthalert D: Amplification of NS3 and NS5A in hepatitis C virus by the multiplex nested polymerase chain reaction. World Acad Sci J 7: 54, 2025.
APA
Sistayanarain, A., & Kunthalert, D. (2025). Amplification of NS3 and NS5A in hepatitis C virus by the multiplex nested polymerase chain reaction. World Academy of Sciences Journal, 7, 54. https://doi.org/10.3892/wasj.2025.342
MLA
Sistayanarain, A., Kunthalert, D."Amplification of NS3 and NS5A in hepatitis C virus by the multiplex nested polymerase chain reaction". World Academy of Sciences Journal 7.4 (2025): 54.
Chicago
Sistayanarain, A., Kunthalert, D."Amplification of NS3 and NS5A in hepatitis C virus by the multiplex nested polymerase chain reaction". World Academy of Sciences Journal 7, no. 4 (2025): 54. https://doi.org/10.3892/wasj.2025.342
Copy and paste a formatted citation
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Spandidos Publications style
Sistayanarain A and Kunthalert D: Amplification of NS3 and NS5A in hepatitis C virus by the multiplex nested polymerase chain reaction. World Acad Sci J 7: 54, 2025.
APA
Sistayanarain, A., & Kunthalert, D. (2025). Amplification of NS3 and NS5A in hepatitis C virus by the multiplex nested polymerase chain reaction. World Academy of Sciences Journal, 7, 54. https://doi.org/10.3892/wasj.2025.342
MLA
Sistayanarain, A., Kunthalert, D."Amplification of NS3 and NS5A in hepatitis C virus by the multiplex nested polymerase chain reaction". World Academy of Sciences Journal 7.4 (2025): 54.
Chicago
Sistayanarain, A., Kunthalert, D."Amplification of NS3 and NS5A in hepatitis C virus by the multiplex nested polymerase chain reaction". World Academy of Sciences Journal 7, no. 4 (2025): 54. https://doi.org/10.3892/wasj.2025.342
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