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Objective: Drug-resistance mutations are routinely detected using standard Sanger sequencing, which does not detect minor variants with a frequency below 20%. The impact of detecting minor variants generated by ultra-deep sequencing (UDS) on HIV drug-resistance interpretations has not yet been studied. Design: Fifty HIV-1 patients who experienced virological failure were included in this retrospective study. Methods: The HIV-1 UDS protocol allowed the detection and quantification of HIV-1 protease and reverse transcriptase variants related to genotypes A, B, C, F and G. DeepChek-HIV simplified drug-resistance interpretation software was used to compare Sanger sequencing and UDS. Results: The total time required for the UDS protocol was found to be approximately three times longer than Sanger sequencing with equivalent reagent costs. UDS detected allof themutations foundby population sequencingand identifiedadditional resistance variants in all patients. An analysis of drug resistance revealed a total of 643 and 224 clinically relevant mutations by UDS and Sanger sequencing, respectively. Three resistance mutations with more than 20% prevalence were detected solely by UDS: A98S (23%), E138A (21%) and V179I (25%). A significant difference in the drugresistance interpretations for 19 antiretroviral drugs was observed between the UDS and Sanger sequencing methods. Y181C and T215Y were the most frequent mutations associated with interpretation differences. Conclusion: A combination of UDS and DeepChek software for the interpretation of drug resistance results would help clinicians provide suitable treatments. A cut-off of 1% allowed a better characterization of the viral population by identifying additional resistance mutations and improving the drug-resistance interpretation. 2014 Wolters Kluwer Health | Lippincott Williams & Wilkins AIDS 2014, 28:000‐000

Objectives: Many cases of acute autochthonous hepatitic E virus (HEV) hepatitis have been reported in France, mainly from the south. Chronic HEV infection has recently been described in immunosuppressed patients. Although a potential risk of chronicity exists in HIV-infected patients, no survey has been conducted in this population. The aim of this study was to assess the sero-virological prevalence of HEV in French HIV-infected patients. Methods: Two hundred and forty-five HIV-infected patients followed at two Infectious Diseases Departments (one in the south, one in the north) were included from January to March 2009. Sera were collected from all patients and tested using anti-HEV IgG and IgM kits. HEV RNA was systematically amplified in the ORF2 region with an in-house method. The IgG avidity index of all IgG-positive samples was determined. Results: Three of the 133 southern patients showed both anti-HEV IgG and IgM positivities, along with cytolysis and biological cholestasis; HEV RNA was amplified in two of these cases, whereas a low IgG avidity index was observed in all three samples. Twelve of the 130 remaining southern patients (9%) showed anti-HEV IgG positivity. The serological prevalence in the 112 northern patients was 3%, which was significantly lower than in the southern patients (P=0.04). No case of acute hepatitis was reported in the north, whereas the prevalence of patients with biochemical liver abnormalities was similar in both areas (P=0.22). Conclusions: In France, HIV-infected patients are at risk of HEV infection with a serological north-to-south gradient. No case of chronic HEV infection was detected in this study.

Objective: The risk of hepatitis C virus (HCV) transmission in medically assisted procreation (MAP) is debated and some researchers have proposed to exclude MAP for HCV-positive infertile patients. The objectives of this study were to assess the presence of viral RNA in the final preparation of density gradient semen fractions collected from men with chronic HCV and HIV co-infection participating in a MAP program, and to assess whether HIV co-infection influences the rate of the presence of HCV RNA in the semen. Design and methods: The study was based on a cohort of 170 HCV-infected male patients (93 HIV co-infected) participating in a MAP program in a French center. Semen samples were subjected to standard MAP sperm preparation, using density-gradient centrifugation with 40 and 90% layers. All aliquots were tested with a commercially available HCV RNA assay (Roche Monitor), adapted for use with semen after a nucleic HCV RNA extraction modification (Organon Technika). Results: Seminal plasma samples from 19 (11%) patients were HCV RNA positive. The positive HCV viral load in semen was less than 600 IU/ml. None of the 90% fractions from HCV-infected patients were HCV RNA positive. Among the 93 co-infected patients, 10 were positive for HCV RNA in semen and three were HIV/HCV RNA positive in semen. Conclusions: Although HCV RNA was found in the semen of 11% of patients, no purified sperm fraction, or spermatozoa used in MAP were HCV RNA positive. The 90% purified sperm fraction discards the virus and must be used with care in MAP.