A Novel Simple Assay System to Quantify the Percent HCV-RNA Levels of NS5A Y93H Mutant Strains and Y93 Wild-Type Strains Relative to the Total HCV-RNA Levels to Determine the Indication for Antiviral Therapy with NS5A Inhibitors

作者：Uchida Yoshihito; Kouyama Jun ichi; Naiki Kayoko; Mochida Satoshi^*

来源：PLos One, 2014, 9(11): e112647.

DOI：10.1371/journal.pone.0112647

Aim: Oral treatment with asunaprevir and daclatasvir has been reported to yield a SVR ratio of 80% in patients with genotype 1b HCV infection, however, treatment failure has been reported, especially in patients with HCV strains showing the NS5A-Y93H mutation at baseline. An assay system to detect such strains was established to facilitate selection of appropriate candidates for this antiviral therapy. %26lt;br%26gt;Methods: Primer sets and 2 types of cycling probe mixtures were designed, and real-time PCR was performed with HCV-RNA purified from 332 patients with genotype 1b HCV infection, and the results were compared with those obtained by direct sequencing. %26lt;br%26gt;Results: Both the wild-type and mutant strains were quantified, with a threshold of 4.0 Log copies/mL, in 295 of the 332 patients (88.9%), and the percentage of the mutant strains relative to the total HCV-RNA level in the serum was calculated. The percentage was 0% in 237 patients (80.3%) and 100% in 23 patients (7.8%), identical to the results of direct sequencing. Both wild-type and mutant strains were detected in the remaining 35 patients (11.9%), at levels between 1% and 99%, despite the mutant strains having been undetectable by direct sequencing in 11 patients with percentages of these strains of less than 25%. %26lt;br%26gt;Conclusion: A novel assay system to quantify the percent RNA of Y93H mutant strains relative to the total HCV-RNA level was established. This system may be useful to determine the indication for treatment with NA5A inhibitors in patients with HCV.

• 出版日期2014-11-14