Evolution of the HCV viral population from a patient with S282T detected at relapse after sofosbuvir monotherapy.

Clinical phase II/III studies of the nucleotide analogue HCV NS5B inhibitor sofosbuvir (SOF) have demonstrated high efficacy in HCV-infected patients in combination therapy. To date, resistance to SOF (S282T in NS5B) has rarely been detected in patients. In this study, we investigated the evolution of S282T viral variants detected in one HCV genotype 2b-infected patient who relapsed following 12 weeks of SOF monotherapy. Deep sequencing of the NS5B gene was performed on longitudinal plasma samples at baseline, days 2 and 3 on SOF, and longitudinal samples post-SOF treatment through week 48. Intrapatient HCV evolution was analysed by maximum-likelihood phylogenetic analysis. Deep sequencing analysis revealed a low level pre-existence of S282T at 0.05% of viral sequences (4/7755 reads) at baseline and 0.03% (6/23 415 reads) at day 2 on SOF. Viral relapse was detected at week 4 post-treatment where 99.8% of the viral population harboured S282T. Follow-up analysis determined that S282T levels diminished post-treatment reaching undetectable levels 24-48 weeks post-SOF. Phylogenetic analysis together with the persistence of unique post-treatment mutations in all post-SOF samples suggested that growth of wild type resulted from reversion of the S282T mutant to a wild type and not outgrowth of the baseline wild-type population. Our data suggest that a very low level of pre-existing S282T at baseline in this patient was enriched and transiently detected following SOF monotherapy. Despite relapse with drug resistance to SOF, this patient was successfully retreated with SOF plus ribavirin for 12 weeks and is now cured from HCV infection.

Bacterial diversity in surgical site infections: not just aerobic cocci any more.

OBJECTIVE: To evaluate the microbial diversity in chronic surgical site infections (SSIs). METHOD: Bacterial populations in 23 chronic SSIs were identified using bacterial tag-encoded FLX amplicon pyrosequencing (bTEFAP),which is an universal bacterial identification method.These results were then validated using quantitative polymerase chain reaction (qPCR). RESULTS: bTEFAP identified two previously uncharacterised Bacteroidales in all of the SSIs and showed that it was the predominant population in the majority of these chronic wounds. Other bacteria identified included Corynebacterium spp., Peptoniphilus spp., Staphylococcus spp., Staphylococcus aureus, Serratia marcescens, Prevotella spp. and Pseudomonas aeruginosa. Rarefaction analysis of the data indicated that, on average, six genera occurred in any given SSI, suggesting that such infections are multispecies. On average, over 60% of the bacteria evaluated in the SSIs were anaerobic bacilli. The previous literature indicates that aerobic cocci predominate in such wounds. CONCLUSION: This modern molecular survey indicates that our previous understanding of which bacteria cause SSIs may be faulty. The high prevalence of anaerobic bacilli and the overwhelming predominance of two previously uncharacterised Bacteroidales suggest that such bacteria may be a leading contributor to such infections. Further research on the identification and treatment of such bacteria are warranted.