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Identification of flaviviruses and phleboviruses from insects in southwest of Spain
S6
Abstracts / Journal of Clinical Virology 70 (2015) S1–S126
Abstract No: 1700
Abstract No: 1720
Presentation at ESCV 2015: Oral 12 Identification of flaviviruses and phleboviruses from insects in southwest of Spain
Presentation at ESCV 2015: Oral 13 In UTERO treatment of cytomegalovirus congenital infection with valacyclovir (CYMEVAL II) NCT01651585
F. Del Ventura-Villarroel 1,∗ , S. Ruiz 2 , L. Herrero 1 , J. Figuerola 3 , L. Hernández 1 , J. Moreno 2 , R. Soriguer 3 , M.P. Sánchez-Seco 1 , A. Vázquez 1 1
Institute of Health Carlos III, Majadahonda, Spain Servicio Control Mosquitos, Huelva, Spain 3 EBD-CSIC, Seville, Spain
M. Leruez-Ville 1,∗ , I. Ghout 2 , J.-F. Magny 1 , S. Couderc 3 , F. Jacquemard 3 , Y. Ville 1 1
Hopital Necker-Enfants-Malades, France Hopital Ambroise Pare, France 3 Hopital Intercommunal de Poissy, France
2
2
Background: In Spain, the flavivirus West Nile (WNV) and the phlebovirus Toscana (TOSV) are authoctonous. They cause neurological disease in humans and are transmitted by mosquitoes (Culex sp.) or sandflies (Phlebotomus sp.), respectively. It seems that these arboviruses are circulating in several areas from Andalucia. In the province of Cádiz in 2010, human infections due to both viruses have been diagnosed during the first WNV outbreak occurred in humans in our country. Moreover, WNV lineage 1 has been detected in Cx. perexiguus from Huelva in 2008 and a new lineage was described in 2006 from Cx. pipiens in the same area. TOSV was detected in Phlebotomus sp. in Granada in 2004. In addition to these viruses, whose pathogenicity has already been demonstrated, other related viruses have been detected in our territory, such as Granada (GRV) and Usutu (USUV), for which its public health importance has not been determined yet. The proposed objective in this work was to study the presence of arboviruses in vectors from several provinces of Southwestern Spain. Methods: Insects belonging to the genera Phlebotomus or Culex were captured in several Andalusian provinces (Huelva, Sevilla and Cádiz) in 2013. Insects were pooled by species, sex, collection site, and date. The arboviral screening was performed with two generics nested RT-PCR to detect flavivirus or phlebovirus genome. A total of 2628 insects (190 pools), belonging to the Psychodidae and Culicidae families, were analyzed. Viral RNA was extracted using the kit QIAamp Viral RNA extraction (Qiagen). Amplification products were sequenced, and sequences were compared with all published sequences recovered from GenBank. They were aligned using CLUSTAL X, and phylogenetic analyses were performed using the MEGA package (version 5.0). Results: Both, flaviviruses and phleboviruses were detected in this work. TOSV was detected in Phlebotomus sp. from Cadiz, and its sequence showed a level of homology of 98% with other sequences of TOSV detected previously in Spain. WNV was detected in three pools of Cx. perexiguus from Sevilla, and these sequences were similar to the WNV sequence detected in 2008 in Cx. perexiguus from Huelva. Moreover, sequences related to the phlebovirus GRV and to the insect flavivirus SCxFV, were detected. Conclusion: In this work, we show the presence of WNV and TOSV in their vectors from some areas in Spain for the first time. These results confirm that these viruses are circulating in a wider area in the Southwest of Spain.
Background: No trial has ever tested the efficacy of any antiviral to treat CMV infected fetuses. Valacyclovir was chosen because of its tolerance in pregnancy and its efficacy in prophylaxis treatment of CMV infection in immunocompromised adults. A first double blind RCT of valacyclovir against placebo failed to complete due to lack of recruitment and relatively easy availability of valacyclovir outside the study. Here, we conducted a phase-2 trial to evaluate the effect of valacyclovir in symptomatic fetuses using the Optimal Two-Stage Simon’ design. Method: The trial was prospective, multicenter, nonrandomized with a single group assignment, all women receiving: valacyclovir (2 g × 4 per day) until delivery. The number of subjects calculated by the optimal 2 stages Simon’ design (based on p0 = non acceptable proportion of asymptomatic infants <60%, p1 = acceptable proportion of asymptomatic infants “d80%, 5% risk and 20% type) required that 7 of the first 11 (70%) cases (first step) and no less than 30 of the overall 43 cases (70%) (second step) should be asymptomatic at birth to show an effect of valacyclovir. Inclusion criteria: Fetal infection (positive CMV PCR in the amniotic fluid), and at least one extra cerebral ultrasound (US) abnormality, and/or one isolated mild cerebral US abnormalities: and/or biological abnormalities: fetal viremia >3000 copies/ml, platelet <100 000/mm3 . The primary end point was to observe a reduction of unfavorable outcomes (symptomatic children at birth) and a reduction in the number of termination of pregnancy for fetal anomalies. Results: At interim analysis 8 of the first 11 cases were asymptomatic (72%) allowing continuing to step 2. Altogether 41 women were included (43 fetuses) at a median 25.9 weeks gestation IQ: 24–31. Median duration of treatment was 89 days IQ: 41–102. Safety was not an issue with only 2 women presenting with mild side effect (headaches) and no liver or renal toxicities. The outcome was: 34 asymptomatic neonates (79.5%), 2 TOP and 7 symptomatic neonates. Conclusion: This first trial evaluating efficacy and safety of valacyclovir to treat symptomatic CMV infected fetuses was successful. This opens the field to trials that could test new antiviral with more potent effect on CMV randomized against ValACV avoiding an unacceptable placebo arm.
http://dx.doi.org/10.1016/j.jcv.2015.07.022
http://dx.doi.org/10.1016/j.jcv.2015.07.023
Abstracts / Journal of Clinical Virology 70 (2015) S1–S126
Abstract No: 1700
Abstract No: 1720
Presentation at ESCV 2015: Oral 12 Identification of flaviviruses and phleboviruses from insects in southwest of Spain
Presentation at ESCV 2015: Oral 13 In UTERO treatment of cytomegalovirus congenital infection with valacyclovir (CYMEVAL II) NCT01651585
F. Del Ventura-Villarroel 1,∗ , S. Ruiz 2 , L. Herrero 1 , J. Figuerola 3 , L. Hernández 1 , J. Moreno 2 , R. Soriguer 3 , M.P. Sánchez-Seco 1 , A. Vázquez 1 1
Institute of Health Carlos III, Majadahonda, Spain Servicio Control Mosquitos, Huelva, Spain 3 EBD-CSIC, Seville, Spain
M. Leruez-Ville 1,∗ , I. Ghout 2 , J.-F. Magny 1 , S. Couderc 3 , F. Jacquemard 3 , Y. Ville 1 1
Hopital Necker-Enfants-Malades, France Hopital Ambroise Pare, France 3 Hopital Intercommunal de Poissy, France
2
2
Background: In Spain, the flavivirus West Nile (WNV) and the phlebovirus Toscana (TOSV) are authoctonous. They cause neurological disease in humans and are transmitted by mosquitoes (Culex sp.) or sandflies (Phlebotomus sp.), respectively. It seems that these arboviruses are circulating in several areas from Andalucia. In the province of Cádiz in 2010, human infections due to both viruses have been diagnosed during the first WNV outbreak occurred in humans in our country. Moreover, WNV lineage 1 has been detected in Cx. perexiguus from Huelva in 2008 and a new lineage was described in 2006 from Cx. pipiens in the same area. TOSV was detected in Phlebotomus sp. in Granada in 2004. In addition to these viruses, whose pathogenicity has already been demonstrated, other related viruses have been detected in our territory, such as Granada (GRV) and Usutu (USUV), for which its public health importance has not been determined yet. The proposed objective in this work was to study the presence of arboviruses in vectors from several provinces of Southwestern Spain. Methods: Insects belonging to the genera Phlebotomus or Culex were captured in several Andalusian provinces (Huelva, Sevilla and Cádiz) in 2013. Insects were pooled by species, sex, collection site, and date. The arboviral screening was performed with two generics nested RT-PCR to detect flavivirus or phlebovirus genome. A total of 2628 insects (190 pools), belonging to the Psychodidae and Culicidae families, were analyzed. Viral RNA was extracted using the kit QIAamp Viral RNA extraction (Qiagen). Amplification products were sequenced, and sequences were compared with all published sequences recovered from GenBank. They were aligned using CLUSTAL X, and phylogenetic analyses were performed using the MEGA package (version 5.0). Results: Both, flaviviruses and phleboviruses were detected in this work. TOSV was detected in Phlebotomus sp. from Cadiz, and its sequence showed a level of homology of 98% with other sequences of TOSV detected previously in Spain. WNV was detected in three pools of Cx. perexiguus from Sevilla, and these sequences were similar to the WNV sequence detected in 2008 in Cx. perexiguus from Huelva. Moreover, sequences related to the phlebovirus GRV and to the insect flavivirus SCxFV, were detected. Conclusion: In this work, we show the presence of WNV and TOSV in their vectors from some areas in Spain for the first time. These results confirm that these viruses are circulating in a wider area in the Southwest of Spain.
Background: No trial has ever tested the efficacy of any antiviral to treat CMV infected fetuses. Valacyclovir was chosen because of its tolerance in pregnancy and its efficacy in prophylaxis treatment of CMV infection in immunocompromised adults. A first double blind RCT of valacyclovir against placebo failed to complete due to lack of recruitment and relatively easy availability of valacyclovir outside the study. Here, we conducted a phase-2 trial to evaluate the effect of valacyclovir in symptomatic fetuses using the Optimal Two-Stage Simon’ design. Method: The trial was prospective, multicenter, nonrandomized with a single group assignment, all women receiving: valacyclovir (2 g × 4 per day) until delivery. The number of subjects calculated by the optimal 2 stages Simon’ design (based on p0 = non acceptable proportion of asymptomatic infants <60%, p1 = acceptable proportion of asymptomatic infants “d80%, 5% risk and 20% type) required that 7 of the first 11 (70%) cases (first step) and no less than 30 of the overall 43 cases (70%) (second step) should be asymptomatic at birth to show an effect of valacyclovir. Inclusion criteria: Fetal infection (positive CMV PCR in the amniotic fluid), and at least one extra cerebral ultrasound (US) abnormality, and/or one isolated mild cerebral US abnormalities: and/or biological abnormalities: fetal viremia >3000 copies/ml, platelet <100 000/mm3 . The primary end point was to observe a reduction of unfavorable outcomes (symptomatic children at birth) and a reduction in the number of termination of pregnancy for fetal anomalies. Results: At interim analysis 8 of the first 11 cases were asymptomatic (72%) allowing continuing to step 2. Altogether 41 women were included (43 fetuses) at a median 25.9 weeks gestation IQ: 24–31. Median duration of treatment was 89 days IQ: 41–102. Safety was not an issue with only 2 women presenting with mild side effect (headaches) and no liver or renal toxicities. The outcome was: 34 asymptomatic neonates (79.5%), 2 TOP and 7 symptomatic neonates. Conclusion: This first trial evaluating efficacy and safety of valacyclovir to treat symptomatic CMV infected fetuses was successful. This opens the field to trials that could test new antiviral with more potent effect on CMV randomized against ValACV avoiding an unacceptable placebo arm.
http://dx.doi.org/10.1016/j.jcv.2015.07.022
http://dx.doi.org/10.1016/j.jcv.2015.07.023