NS3 genomic sequencing and phylogenetic analysis as alternative to a commercially available assay to reliably determine hepatitis C virus subtypes 1a and 1b

Objective: To evaluate the use of hepatitis C virus (HCV) NS3 sequencing as alternative to the comercially available Versant HCV 2.0 reverse hybridization line-probe assay (LiPA 2.0) to determine HCV genotype 1 (HCV-1) subtypes. Patients and methods: A cohort of 104 patients infected by HCV-1 according to LiPA 2.0 was analyzed in a cross-sectional study conducted in patients seen from January 2012 to June 2016 at an outpatient clinic in Buenos Aires, Argentina. Results: The samples were included within well supported subtype clades: 64 with HCV-1b and 39 with HCV-1a infection. Twenty of the HCV-1a infected patientes were included in a supported sub-clade “1” and 19 individuals were among the basal sub-clade “2”. LiPA 2.0 failed to subtype HCV-1 in 20 (19.2%) individuals. Subtype classification determined by NS3 direct sequencing showed that 2/18 (11.1%) of the HCV-1a-infected patients as determined by LiPA 2.0 were in fact infected by HCV-1b. Of the HCV-1b-infected according to LiPA 2.0, 10/66 (15.2%) patients showed HCV-1a infection according to NS3 sequencing. Overall misclassification was 14.3% (κ-index for the concordance with NS3 sequencing = 0.635). One (1%) patient was erroneously genotyped as HCV-1 and was revealed as HCV genotype 4 infection. Conclusions: Genomic sequencing of the HCV NS3 region represents an adequate alternative since it provides reliable genetic information. It even distinguishes between HCV-1a clades related to resistance-associated substitutions to HCV protease inhibitors, it provides reliable genetic information for genotyping/subgenotyping and simultaneously allows to determine the presence of resistance-associated substitutions to currently recommended DAAs.Fil: Neukam, Karin. Universidad de Buenos Aires. Facultad de Farmacia y Bioquímica; Argentina. Universidad de Sevilla; España. Hospital Universitario de Valme; EspañaFil: Martínez, Alfredo P.. Centro de Educación Médica e Investigaciones Clínicas “Norberto Quirno”; ArgentinaFil: Culasso, Andrés Carlos Alberto. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Universidad de Buenos Aires. Facultad de Farmacia y Bioquímica; ArgentinaFil: Ridruejo, Ezequiel. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Parque Centenario. CEMIC-CONICET. Centro de Educaciones Médicas e Investigaciones Clínicas "Norberto Quirno". CEMIC-CONICET.; ArgentinaFil: García, Gabriel. Universidad de Buenos Aires. Facultad de Farmacia y Bioquímica; ArgentinaFil: Di Lello, Federico Alejandro. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Universidad de Buenos Aires. Facultad de Farmacia y Bioquímica; Argentin

No evidence of firstly acquired acute hepatitis C virus infection outbreak among HIV-infected patients from Southern Spain: a multicentric retrospective study from 2000-2014

Background: Acute hepatitis C virus (HCV) infection (AHCVI) outbreaks have been described recently within defined areas worldwide among HIV-infected homosexual men. This study aims to describe the cumulative frequency and incidence of firstly acquired AHCVI in an HIV-infected population in Southern Spain. Methods: This is a retrospective study conducted at the Infectious Diseases Units of eight hospitals in Andalusia, Southern Spain. Primary AHC was considered as HCV immunoglobulin G antibody seroconversion. The time of infection was considered the moment between the last negative and the first positive HCV antibody determination. Results: A total of 23 cases of primary AHCVI have been detected from 2000 to 2014. Incidence rates [IR; 95 % confidence interval (CI)] were 0.036 (2.272–0.054) per 100 person-years (py) in the overall population over a follow-up period of 64170 py. Of the 22 (95.7 %) male subjects, 21 (95.5 %) had acquired AHCVI by homosexual contact, the IR (95 % CI) was 0.039 (0.024–0.06) per 100 py in this subpopulation. There was no evidence of an increase of AHCVI IR. The incidence of AHCVI was slightly lower between 2000 and 2004 as compared to 2005–2009 [IR ratio (IRR) of 8.8 (95 % CI: 1.279–378.794; p = 0.01)] but reached a plateau afterwards [IRR between 2010 and 2014 versus 2005–2009: 0.727 (0.286–1.848; p = 0.5)]. The median (Q1-Q3) time between the last negative anti-HCV and the first positive anti-HCV determination was 4.7 (1.9–11.2) months. Peak (Q1-Q3) ALT and total bilirubin values during AHCVI were 496 (291–656) IU/mL and 1.15 (0.9–1.98) mg/dL, respectively. Conclusions: In contrast to what has been reported from other areas, the incidence of primary AHCVI in the HIVinfected population is stable in Southern Spain and there is no evidence of an epidemic, in spite of the high prevalence of HIV/HCV-coinfection in this area.Plan Nacional R + D + I RD12/0017/0012ISCIII-Subdirección General de EvaluaciónFondo Europeo de Desarrollo Regional (FEDER) European Union (EU)Instituto de Salud Carlos III PI15/01124, CP13/00187, Programa-I3SN

High-resolution anoscopy in HIV-infected men: Assessment of the learning curve and factors that improve the performance

To determine the required learning time for high-resolution anoscopy (HRA)-guided biopsy to detect histological high-risk squamous intraepithelial lesions (hHSIL) and to identify factors that impact on the training process. Methods: All HIV-infected, screening-naïve men-who-have-sex-with-men who underwent HRA conducted by one single observer from 2010 to 2017 in a Spanish HIV-outpatient clinic were analysed. Results: Eighty-five (14.7%) of the 581 patients included presented hHSIL. The factors associated with the capacity to detect hHSIL [adjusted odds ratio (aOR), 95% confidence interval (95%CI)] were the presence of cytological HSIL (3.04, 1.78–5.21; p < 0.001), infection with high-risk human papilloma virus (HR-HPV) (2.89, 1.38–6.05; p=0.005), the number of biopsies taken/HRA (aOR: 1.28, 1.07–1.52; p=0.006) and tobacco smoking (1.75; 1.12–2.73; p=0.014). Two events independently augmented the detection rate of hHSIL: one single experienced pathologist interpreted biopsies after 409 HRA (2.80, 1.74–4.48; p=0.035) and the anoscopist underwent an additional training after 536 HRA (2.57, 1.07–6.16; p=0.035). A learning process could be observed throughout the whole study with stable HR-HPV prevalence. Conclusion: The data support the growing evidence that the proposed training volume of 50–200 performances is underestimated. Extensive training of both anoscopist and pathologist is warranted and the development of tools to support the diagnostic performance may be considered.Plan Nacional R+D+I y Red de Investigación en SIDA RD16/0025/0020-ISCIII-FEDMiguel Servet research grant, Instituto de Salud Carlos III CPII18/0003

Reassessment of genotype 1 hepatitis c virus subtype misclassification by LiPA 2.0: implications for direct-acting antiviral treatment

The accuracy of LiPA 2.0 for hepatitis C virus 1 (HCV-1) subtype classification was analyzed. LiPA 2.0 genotype results from 101 HCV-1-infected patients were compared to genotype findings determined by direct core sequencing. Eleven (11%) samples were misclassified. Given the influence of the HCV-1-subtype in the anti-HCV therapy response, an alternative classification method is warranted.Fil: Guelfo, Javier R.. Hospital Universitario de Valme. Unidad de Enfermedades Infecciosas y Microbiología; EspañaFil: Macias, Juan. Hospital Universitario de Valme. Unidad de Enfermedades Infecciosas y Microbiología; EspañaFil: Neukam, Karin. Hospital Universitario de Valme. Unidad de Enfermedades Infecciosas y Microbiología; EspañaFil: Di Lello, Federico Alejandro. Hospital Universitario de Valme. Unidad de Enfermedades Infecciosas y Microbiología; España. Universidad de Buenos Aires. Facultad de Farmacia y Bioquímica; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; ArgentinaFil: Mira José Antonio. Hospital Universitario de Valme. Unidad de Enfermedades Infecciosas y Microbiología; EspañaFil: Merchante, Nicolás. Hospital Universitario de Valme. Unidad de Enfermedades Infecciosas y Microbiología; EspañaFil: Mancebo, María. Hospital Universitario de Valme. Unidad de Enfermedades Infecciosas y Microbiología; EspañaFil: Nuñez Torres, Rocío. Hospital Universitario de Valme. Unidad de Enfermedades Infecciosas y Microbiología; EspañaFil: Pineda, Juan A.. Hospital Universitario de Valme. Unidad de Enfermedades Infecciosas y Microbiología; EspañaFil: Real. Luis M.. Hospital Universitario de Valme. Unidad de Enfermedades Infecciosas y Microbiología; Españ

Heterologous Gam-COVID-Vac (Sputnik V)/mRNA-1273 (Moderna) vaccination: Author's response

We appreciate the observation from the authors Rujittika Mungmunpuntipantip and Viroj Wiwanitkit [1] regarding our publication [2]. Certainly, there is a vast number of factors that may affect immunological parameters from a greater to a lesser extent. We agree this is an important aspect, so the analysis of the immunological parameters assessed in our work has considered the broadly established factors that may have potentially impacted the immune response in our setting. Although elevated anti-S-RBD immunoglobulin (IgG) values after asymptomatic COVID-19 cannot be excluded, both groups of our comparative study would have been affected similarly; therefore, an impact on the conclusions would unlikely be altered. Moreover, the detection of the nucleoprotein IgG antibody to distinguish vaccine-induced seropositivity from natural infection is limited due to its significant decrease after 120 days postinfection period [3]. Consequently, we felt that the analytical approach initially performed in our research was accurate. Finally, the immune response variations according to different boost vaccination schemes observed in our study were also registered in a subset of patients receiving dialysis [4] and confirmed by a recently published study that assessed the humoral response to several vaccination schemes, including the homologous prime-boost vaccination with Gam-COVID-Vac and the heterologous combination with mRNA-1273 (Moderna) vaccine [5].Peer reviewe

Assessment of the humoral response to the homologous Gam-COVID-Vac (Sputnik V) or heterologous Sputnik V/mRNA-1273 (Moderna) vaccination against SARS-CoV-2 in dialysis patients

Background and Aim Dialysis patients are a high-risk population and have a reduced immune response to vaccination against SARS-CoV-2. The aim of this study was to assess the humoral response to homologous Gam-COVID-Vac (Sputnik V) and heterologous Sputnik V/mRNA-1273 (Moderna) vaccination in dialysis patients. The vaccination scheme depended on dose availability and the prioritization of risk populations as established by the Argentine Ministry of Health. Methods Previous COVID-19 infection was determined in symptomatic patients. Binding IgG antibodies against the spike (S) receptor-binding domain (RBD) of SARS-CoV-2 (anti-S-RBD) concentration was assessed between 3 and 16 weeks after the boost dose. Anti-S-RBD antibodies were quantified using the Abbott Diagnostics SARS-CoV-2 IgG II Quant chemiluminescent microparticle immunoassay (CMIA) on an Architect i2000 SR and an Alinity I analyzer (Abbott Diagnostics, Abbott Park, Illinois, USA). To standardize the results to WHO binding antibody units (BAU), a correction factor for Abbott arbitrary units (AU) was applied where 1 BAU/mL equals 0.142 AU, as previously established by Abbott with the WHO international standard NIBSC 20–136. Following the manufacturer’s recommendations, samples were considered reactive for anti-S-RBD when titers were above 50 AU/mL (7.2 BAU/mL). An 80% protective effect (PROT-80) against symptomatic SARS-CoV-2 infection was assumed when anti-S-RBD titers were 506 BAU/ml or higher. Charlson Comorbidity Index (CCI) score was classified as mild = 1–2, moderate = 3–4, and severe ≥ 5. Side effects were evaluated until day 7 by patients´ self-reported questionnaire. Results One hundred seven participants were enrolled [n = 84 homologous (SpV/SpV), nn 23 heterologous (SpV/Mod)]. Median (IQR) age was 64 (50–75) years old and 79 (73.8%) were male. Additionally, 19 (22.6%) of the SpV/SpV and 4 (17.4%) of the SpV/Mod group had a prior confirmed SARS-CoV-2 infection (p = 0.589). In the overall population, 103 patients reached seroconversion (96.3%). Anti-S-RBD IgG median titers (IQR) were higher in the heterologous [1222 (288–5680) BAU/mL] than in the homologous scheme [447 (100–1551) BAU/mL], p = 0.022. In a linear model adjusted for age, gender, days from first vaccination to boost dose and days from the boost dose to the anti-S-RBD IgG determination, previous SARS-COV-2 infection (B: 2062.2; CI95: 1231.8–2892.6; p < 0.001), and SpV/Mod vaccination scheme (B: 1294.6; CI95: 435.58–2147.6; p = 0.003) were independently associated with anti-S-RBD levels. Finally, a higher frequency of adverse effects was associated with the heterologous scheme, although they were well tolerated by all individuals. Conclusions The present study provides evidence that the homologous SpV/SpV and heterologous SpV/Mod schemes showed good efficacy and safety in patients on chronic dialysis. These results could be useful for designing future vaccination strategies, especially aimed at this risk group.FAD is a member of the National Research Council (CONICET) Research Career Program. K.N. is the recipient of a Miguel Servet contract by the Instituto de Salud Carlos III (grant number CPII18/00033). We would like to thank Mrs. Silvina Heisecke, from CEMIC‐CONICET, for the copyediting of the manuscriptPeer reviewe

Humoral response to the Sputnik V and Sputnik V/Moderna in dialysis

Introduction: The humoral response to vaccines is the most used tool to evaluate the protection against SARS-CoV-2 infection. Dialysis patients are a high-risk population and have a reduced immune response to vaccination. Objective: To assess the humoral response to homologous Gam-COVID-Vac (Sputnik V) and heterologous Sputnik V/mRNA-1273 (Moderna) vaccination in dialysis patients. Methods: SARS-CoV-2 anti-spike IgG (RBD) concentration was estimated 3-16 weeks after complete vaccination. Reactogenicity was evaluated until day 7 by patients self-reported side events. Results: 107 participants were enrolled [n=84 homologous (SpV/SpV), n=23 heterologous (SpV/Mod)]. Median (IQR) age was 64 (50-75) years old and 79 (73.8%) were male. Additionally, 19 (22.6%) of the SpV/SpV and 4 (17.4%) of the SpV/Mod group had a prior confirmed SARS-CoV-2 infection (p=0.589). In the overall population, 103 patients reached seroconversion (96.3%). Anti-S-RBD IgG median titers (IQR) were higher in the heterologous [1222 (288-5680) BAU/mL] than in the homologous scheme [447 (100-1551) BAU/mL], p=0.022. In a linear model adjusted for age and gender, previous SARS-COV-2 infection (B: 1944.3; CI95: 1136.2-2753.4; p<0.001), and SpV/Mod vaccination scheme (B: 1241.5; CI95: 420.39-2062.6; p=0.003) were independently associated with anti-S-RBD levels. Finally, a higher frequency of adverse effects was associated with the heterologous scheme, although they were well tolerated by all individuals. Conclusion: The present study provides evidence that the homologous SpV/SpV and heterologous SpV/Mod schemes showed good efficacy and safety under dialysis conditions. These results could be useful for future vaccination strategies, especially aimed at this risk group.FAD is a member of the National Research Council (CONICET) Research Career Program. K.N. is the recipient of a Miguel Servet contract by the Instituto de Salud Carlos III (grant number CPII18/00033). We would like to thank Mrs. Silvina Heisecke, from CEMIC‐CONICET, for the copyediting of the manuscriptN

Heterologous gam-COVID-vac (sputnik V)/mRNA-1273 (moderna) vaccination induces a stronger humoral response than homologous sputnik V in a real-world data analysis

Objectives To compare the homologous prime-boost vaccination scheme of Gam-COVID-Vac (Sputnik V (SpV)) to its heterologous combination with mRNA-1273 (Moderna (Mod)) vaccine. Methods SARS-CoV-2 anti-spike (S)-receptor binding domain (RBD) IgG concentration was assessed three to seven weeks after complete vaccination. Reactogenicity was evaluated by declared side events and medical assistance required until day 7 post boost. Results Of 190 participants enrolled, 105 received homologous SpV/SpV and the remaining heterologous SpV/Mod vaccination scheme, respectively. Median (interquartile range (IQR)) age was 54 (37–63) years, 132 out of 190 (69.5%) were female, and 46 out of 190 (24.2%) individuals had a prior confirmed COVID-19. Anti-S-RBD IgG median (IQR) titers were significantly higher for SpV/Mod (2511 (1476–3992) binding antibody units (BAU)/mL) than for SpV/SpV (582 (209–1609) BAU/mL; p < 0.001] vaccination scheme. In a linear model adjusted for age, gender, time to the serological assay, and time between doses, SpV/Mod (4.154 (6.585–615.554); p < 0.001] and prior COVID (3.732 (8.641–202.010); p < 0.001) were independently associated with higher anti-S-RBD IgG values. A higher frequency of mild and moderate adverse effects was associated with the heterologous scheme (20 of 85 (23.5%) vs. 13 of 105 (12.4%); p = 0.043 and 27 of 85 (31.8%) vs. 14 of 105 (13.3%); p = 0.002), respectively, although it was well tolerated by all individuals and no medical assistance was required. Discussion The heterologous SpV/Mod combination against SARS-CoV-2 is well tolerated and significantly increases humoral immune response as compared to the homologous SpV/SpV immunization.Matías J. Pereson, Patricia Baré, María Noel Badano and Federico A. Di Lello are members of the National Research Council (CONICET) Research Career Program. Karin Neukam is the recipient of a Miguel Servet contract by the Instituto de Salud Carlos III (grant number CPII18/00033).Peer reviewe

Heterologous Gam-COVID-Vac (Sputnik V) / mRNA-1273 (Moderna) vaccination induces a stronger humoral response than homologous Sputnik V in a real-world data analysis

Introduction Growing data are demonstrating safety and immunogenicity of heterologous vaccination schemes against severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2) infection. This strategy opens up the possibility of a shorter path towards the end of the pandemic. Objective To compare the homologous prime-boost vaccination scheme of Gam-COVID-Vac (Sputnik V, SpV) to its heterologous combination with mRNA-1273 (Moderna, Mod) vaccine. Methods SARS-CoV-2 anti-spike (S)-receptor binding domain (RBD) IgG concentration was assessed three to seven weeks after complete vaccination. Reactogenicity was evaluated by declared side events and medical assistance required until day 7 post-boost. Results Of 190 participants enrolled, 105 received homologous SpV/SpV and the remaining heterologous SpV/Mod vaccination scheme, respectively. Median (interquartile range, IQR) age was 54 (37-63) years, 132 (69.5%) were female and 46 (24.2%) individuals had a prior confirmed COVID-19. Anti-S-RBD IgG median (IQR) titers were significantly higher for SpV/Mod [2511 (1476-3992) BAU/mL] than for SpV/SpV [582 (209-1609) BAU/mL, p<0.001] vaccination scheme. In a linear model adjusted for age, gender, time to the serological assay and time between doses, SpV/Mod [4.154 (6.585-615.554), p<0.001] and prior COVID [3.732 (8.641-202.010), p<0.001] were independently associated with higher anti-S-RBD IgG values. A higher frequency of mild-moderate adverse effects was associated with the heterologous scheme, although it was well tolerated by all individuals and no medical assistance was required. Conclusion The heterologous SpV/Mod combination against SARS-CoV-2 is well tolerated and significantly increases humoral immune response as compared to the homologous SpV/SpV immunization.N

No evidence of firstly acquired acute hepatitis C virus infection outbreak among HIV-infected patients from Southern Spain: a multicentric retrospective study from 2000-2014

[Background] Acute hepatitis C virus (HCV) infection (AHCVI) outbreaks have been described recently within defined areas worldwide among HIV-infected homosexual men. This study aims to describe the cumulative frequency and incidence of firstly acquired AHCVI in an HIV-infected population in Southern Spain.[Methods] This is a retrospective study conducted at the Infectious Diseases Units of eight hospitals in Andalusia, Southern Spain. Primary AHC was considered as HCV immunoglobulin G antibody seroconversion. The time of infection was considered the moment between the last negative and the first positive HCV antibody determination.[Results] A total of 23 cases of primary AHCVI have been detected from 2000 to 2014. Incidence rates [IR; 95 % confidence interval (CI)] were 0.036 (2.272–0.054) per 100 person-years (py) in the overall population over a follow-up period of 64170 py. Of the 22 (95.7 %) male subjects, 21 (95.5 %) had acquired AHCVI by homosexual contact, the IR (95 % CI) was 0.039 (0.024–0.06) per 100 py in this subpopulation. There was no evidence of an increase of AHCVI IR. The incidence of AHCVI was slightly lower between 2000 and 2004 as compared to 2005–2009 [IR ratio (IRR) of 8.8 (95 % CI: 1.279–378.794; p = 0.01)] but reached a plateau afterwards [IRR between 2010 and 2014 versus 2005–2009: 0.727 (0.286–1.848; p = 0.5)]. The median (Q1-Q3) time between the last negative anti-HCV and the first positive anti-HCV determination was 4.7 (1.9–11.2) months. Peak (Q1-Q3) ALT and total bilirubin values during AHCVI were 496 (291–656) IU/mL and 1.15 (0.9–1.98) mg/dL, respectively.[Conclusions] In contrast to what has been reported from other areas, the incidence of primary AHCVI in the HIV-infected population is stable in Southern Spain and there is no evidence of an epidemic, in spite of the high prevalence of HIV/HCV-coinfection in this area.This work has been partially funded by the RD12/0017/0012 project as part of the Plan Nacional R + D + I and cofinanced by ISCIII-Subdirección General de Evaluación, the Fondo Europeo de Desarrollo Regional (FEDER), and the Instituto de Salud Carlos III (grant number PI15/01124). K.N. is the recipient of a Miguel Servet research grant from the Instituto de Salud Carlos III (grant number CP13/00187). J.M. is the recipient of a grant from the Servicio Andaluz de Salud de la Junta de Andalucía (grant number B-0037). J.A.P. is recipient of an intensification grant from the Instituto de Salud Carlos III (grant number Programa-I3SNS).Peer reviewe